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櫻谷英治

徳島大学

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2024年11月22日更新

職名: 教授
電話: 研究者総覧に該当データはありませんでした。
電子メール: sakuradani.eiji@tokushima-u.ac.jp
学歴: 研究者総覧に該当データはありませんでした。
学位: 博士(農学)
職歴・経歴: 2003/6: 京都大学大学院農学研究科助手
2007/4: 京都大学大学院農学研究科助教
2010/10: カナダブリティッシュコロンビア大学植物学部研究員
2014/5: 徳島大学大学院ソシオテクノサイエンス研究部教授
2016/4: 徳島大学生物資源産業学部教授

専門分野・研究分野: ライフサイエンス (Life sciences) [応用生物化学 (Applied biochemistry)]

研究者総覧で最新データを確認する

2024年11月22日更新

専門分野・研究分野: ライフサイエンス (Life sciences) [応用生物化学 (Applied biochemistry)]
担当経験のある授業科目: キャリアパス (学部)
フードサイエンス (学部)
卒業研究 (学部)
基礎食品化学 (学部)
微生物分子論 (大学院)
応用微生物学 (学部)
応用微生物学Ⅰ (学部)
応用微生物学Ⅱ (学部)
応用微生物学特論 (大学院)
応用発酵学 (学部)
生物資源学研究 (大学院)
生物資源産業学B (学部)
生物資源産業学実習 (学部)
生物資源産業学専門英語 (学部)
生物資源産業学概論 (学部)
発酵醸造学 (学部)
英語論文講読 (学部)
英語論文講読Ⅰ (学部)
英語論文講読Ⅱ (学部)
食と健康概論 (学部)
食料生物科学特別実習 (大学院)
食料生物科学特別演習 (大学院)
食料生物科学特別研究 (大学院)
食料生物科学特別講義 (大学院)
食料科学基礎実習 (学部)
食料科学実習A (学部)
食料科学実習B (学部)
食料科学実習C (学部)
食料科学実習Ⅰ (学部)
食料科学実習Ⅱ (学部)
食料科学概論 (学部)
指導経験: 29人 (学士), 27人 (修士), 2人 (博士)

研究者総覧で最新データを確認する

2024年11月22日更新

専門分野・研究分野: ライフサイエンス (Life sciences) [応用生物化学 (Applied biochemistry)]

研究テーマ: 研究者総覧に該当データはありませんでした。

著書

安藤晃規, 奥田知生, 菊川寛史, 阪本鷹行, 櫻谷英治, 小川順 :
油糧微生物による種々のω3脂肪酸含有油脂の発酵生産,
株式会社シーエムシー出版, 2021年4月. 菊川寛史, 奥田知生, 安藤晃規, 櫻谷英治, 小川順 :
Mortierella alpina 1S-4の分子育種による種々のPUFAの生産,
日本生物工学会, 2020年.

(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1520009407267856128

(CiNii: 1520009407267856128) 櫻谷英治, 阪本鷹行 :
廃グリセロールを利用した油脂発酵生産,
三恵社, 2016年11月. jun Ogawa, Eiji Sakuradani, Shigenobu Kishino, Akinori Ando, Kenzo Yokozeki and Sakayu Shimizu :
Microbial Production of Functional Polyunsaturated Fatty Acids and Their Derivatives,
2014.

(出版サイトへのリンク): ● Publication site (DOI): 10.1007/978-4-431-54607-8_18
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1007/978-4-431-54607-8_18

(DOI: 10.1007/978-4-431-54607-8_18) 安藤晃規, 櫻谷英治, 小川順 :
油糧糸状菌Mortierella alpinaによるEPAの常温発酵生産,
2013年. 小川順, 櫻谷英治, 岸野重信, 安藤晃規, 清水昌 :
バイオテクノロジーシリーズ「微生物を活用した新世代の有用物質生産技術」第Ⅲ編穴澤秀治監修 " 第7章高度不飽和脂肪酸・共役脂肪酸含有油脂の微生物生産",
株式会社シーエムシー出版, 東京, 2012年9月. Eiji Sakuradani, Akinori Ando, Jun Ogawa and Sakayu Shimizu :
Single Cell Oils: Microbial and Algal Oils (eds. by Z. Cohen and C. Ratledge) "Arachidonic Acid-Producing Mortierella alpina: Creation of Mutants, Isolation of the Related Enzyme Genes, and Molecular Breeding",
American Oil Chemists' Society, Urbana, May 2010. 小川順, 櫻谷英治, 岸野重信, 安藤晃規, 清水昌 :
酵素利用技術大系小宮山眞監修 " 第5編-第1章-第5節「2. 有用脂肪酸の生産」",
株式会社エヌ·ティー·エス, 東京, 2010年4月. 小川順, 岸野重信, 櫻谷英治, 清水昌 :
微生物によるものづくり ―化学法に代わるホワイトバイオテクノロジーの全て― "高度不飽和脂肪酸・共役脂肪酸含有油脂の微生物生産",
株式会社シーエムシー出版, 東京, 2008年6月. 小川順, 岸野重信, 櫻谷英治, 横関健三, 清水昌 :
栄養学研究の最前線. 日本栄養食糧学会監修 " 微生物機能を活用した食品機能の創出",
株式会社建帛社, 東京, 2008年4月. Eiji Sakuradani, Seiki Takeno, Takahiro Abe, Jun Ogawa and Sakayu Shimizu :
Biocatalysis and Biotechnology for Functional Foods and Industrial Products (eds. by C.T. Hou, J.F. Shew) "Arachidonic acid-producing Mortierella alpina: Molecular breeding of mutants and creation and application of a host-vector system",
CRC Press, New York, Dec. 2006. 櫻谷英治, 清水昌 :
発酵・醸造食品の最新技術と機能性 "脂肪酸発酵による機能性脂質の生産",
株式会社シーエムシー出版, 東京, 2006年7月. Eiji Sakuradani, Seiki Takeno, Takahiro Abe and Sakayu Shimizu :
Single Cell Oils (eds. by Z. Cohen and C. Ratledge) "Arachidonic acid-producing Mortierella alpina: creation of mutants and Molecular breeding",
American Oil Chemists' Society, Urbana, Apr. 2005. 小川順, 櫻谷英治, 清水昌 :
機能性脂質のフロンティア，機能性脂質の分子設計,
株式会社シーエムシー出版, 東京, 2004年12月. Jun Ogawa, Eiji Sakuradani and Sakayu Shimizu :
Lipid Biotechnology (eds. by T.M. Kuo and H.W. Gardner), Production of C20 polyunsaturated fatty acids by an arachidonic acid-producing fungus Mortierella alpina 1S-4 and related strains.,
Mercel Dekker, Inc., New York, 2002.

論文

Hiroshi Kikukawa, Akinori Ando, Asuka Hannya, Mohd Farida Fazli Asras, Tomoyo Okuda, Takaiku Sakamoto, Kiyotaka Y. Hara, Eiji Sakuradani and Jun Ogawa :
Mead acid production by disruption of Δ12-desaturase gene in Mortierella alpina 1S-4,
Journal of Bioscience and Bioengineering, 2023.

(要約): Mead acid (MA; 20:3ω9) is one of the ω9 series of polyunsaturated fatty acids (PUFAs). MA is used to inhibit the inflammation of joints and is applied to the medicinal or health food field. We aimed to construct MA-producing strains with disruption of the Δ12-desaturase gene (Δ12ds) via an efficient gene-targeting system using the lig4-disrupted strain of Mortierella alpina 1S-4 as the host. The transformants showed a unique fatty acid composition that only comprised ω9-PUFAs and saturated fatty acids, while ω6-and ω3-PUFAs were not detected, and the total composition of ω9-PUFAs, including oleic acid (18:1ω9), 18:2ω9, 20:1ω9, 20:2ω9, and MA, was up to 68.4% of the total fatty acids. The MA production in the Δ12ds-disruptant reached 0.10 g/L (8.5%), which exceeded 0.050 g/L (4.6%) in the conventional Δ12ds-defective mutant JT-180.
(キーワード): Mortierella alpina 1S-4 / Gene disruption / Δ12-desaturase / Polyunsaturated fatty acid / Mead acid
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2023.08.001
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 37635046; ● Summary page in Scopus @ Elsevier: 2-s2.0-85169049369

(DOI: 10.1016/j.jbiosc.2023.08.001, PubMed: 37635046, Elsevier: Scopus) Risa Sasaki, Shogo Toda, Takaiku Sakamoto, Eiji Sakuradani and Shinsuke Shigeto :
Simultaneous Imaging and Characterization of Polyunsaturated Fatty Acids, Carotenoids, and Microcrystalline Guanine in Single Aurantiochytrium limacinum Cells with Linear and Nonlinear Raman Microspectroscopy,
The Journal of Physical Chemistry B, Vol.127, No.12, 2708-2718, 2023.

(要約): Thraustochytrids are heterotrophic marine protists known for their high production capacity of various compounds with health benefits, such as polyunsaturated fatty acids and carotenoids. Although much effort has been focused on developing optimal cultivation methods for efficient microbial production, these high-value compounds and their interrelationships are not well understood at the single-cell level. Here we used spontaneous (linear) Raman and multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy to visualize and characterize lipids (e.g., docosahexaenoic acid) and carotenoids (e.g., astaxanthin) accumulated in single living Aurantiochytrium limacinum cells. Spontaneous Raman imaging with the help of multivariate curve resolution alternating least-squares enabled us to make unambiguous assignments of the molecular components we detected and derive their intracellular distributions separately. Near-IR excited CARS imaging yielded the Raman images at least an order of magnitude faster than spontaneous Raman imaging, with suppressed contributions of carotenoids. As the culture time increased from 2 to 5 days, the lipid amount increased by a factor of 7, whereas the carotenoid amount did not change significantly. Furthermore, we observed a highly localized component in A. limacinum cells. This component was found to be mixed crystals of guanine and other purine derivatives. The present study demonstrates the potential of the linear nonlinear Raman hybrid approach that allows for accurate molecular identification and fast imaging in a label-free manner to link information derived from single cells with strategies for mass culture of useful thraustochytrids.
(キーワード): Guanine / Hydrocarbons / Imaging / Lipids / Raman spectroscopy
(徳島大学機関リポジトリ): ● Metadata: 118912
(出版サイトへのリンク): ● Publication site (DOI): 10.1021/acs.jpcb.3c00302
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 36920390; ● Search Scopus @ Elsevier (PMID): 36920390; ● Search Scopus @ Elsevier (DOI): 10.1021/acs.jpcb.3c00302

(徳島大学機関リポジトリ: 118912, DOI: 10.1021/acs.jpcb.3c00302, PubMed: 36920390) Takaiku Sakamoto, Yusuke Ikeda, Naruho Masuda and Eiji Sakuradani :
Ethanol Enhances Astaxanthin Production by Aurantiochytrium sp. O5-1-1,
Journal of Oleo Science, Vol.72, No.4, 441-446, 2023.

(要約): Two-percent ethanol increased the astaxanthin productivity of heterotrophic microalgae Aurantiochytrium sp. O5-1-1 to 2.231 mg/L, 45-fold higher than under ethanol-free condition. Ethanol in the medium decreased at the same rate as spontaneous volatilization, suggesting that it was not a transient signaling factor but a continuous stress on the cells. The triply mutated strain OM3-3 produced 5.075 mg/L astaxanthin under 2% ethanol conditions. Furthermore, the astaxanthin accumulation of the mutant OM3-9 was 0.895 mg/g, which was 150-fold higher than that of strain O5-1-1 in ethanol-free condition. These results are beneficial for the commercial exploitation of carotenoids producing Aurantiochytrium spp.
(キーワード): Aurantiochytrium / astaxanthin / ethanol stress / β-carotene
(徳島大学機関リポジトリ): ● Metadata: 118143
(出版サイトへのリンク): ● Publication site (DOI): 10.5650/jos.ess22308
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 36990751; ● Search Scopus @ Elsevier (PMID): 36990751; ● Search Scopus @ Elsevier (DOI): 10.5650/jos.ess22308

(徳島大学機関リポジトリ: 118143, DOI: 10.5650/jos.ess22308, PubMed: 36990751) Mayu Kikuchi, Keisei Sowa, Michiki Takeuchi, Kasumi Nakagawa, Momoka Matsunaga, Akinori Ando, Kenji Kano, Jun Ogawa and Eiji Sakuradani :
Quantification of leuco-indigo in indigo-dye-fermenting suspension by normal pulse voltammetry,
Journal of Bioscience and Bioengineering, Vol.134, No.1, 84-88, 2022.

(要約): Quantification of leuco-indigo is most important for Aizome, Japanese indigo-dyeing; however, there has been no convenient quantitative method. This study demonstrated that normal pulse voltammetry under quiescent conditions can be used to detect leuco-indigo. As a result of quantification of leuco-indigo in the depth direction in fermenting suspensions, the steady-state concentrations of leuco-indigo showed sigmoidal profiles in the depth direction. The steady state is caused by competitive reactions of microbial reduction of indigo and autoxidation of leuco-indigo by O dissolved from the air interface of the suspension. In addition, we investigated the effects of stirring the suspension and adding some nutrients to the concentration profile. The weakened activity was partially recovered by the addition of ethanol and remarkably recovered by the addition of hipolypepton or glucose. Knowledge is essential for the proper management of indigo-dye-fermenting suspensions.
(キーワード): Coloring Agents / Indigo Carmine / Suspensions
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2022.04.009
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 35597724; ● Search Scopus @ Elsevier (PMID): 35597724; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2022.04.009

(DOI: 10.1016/j.jbiosc.2022.04.009, PubMed: 35597724) Kasumi Nakagawa, Michiki Takeuchi, Manami Tada, Momoka Matsunaga, Masami Kugo, Suzuna Kiyofuji, Mayu Kikuchi, Kazuya Yomota, Takaiku Sakamoto, Kenji Kano, Jun Ogawa and Eiji Sakuradani :
Isolation and characterization of indigo-reducing bacteria and analysis of microbiota from indigo fermentation suspensions,
Bioscience, Biotechnology, and Biochemistry, Vol.86, No.2, 273-281, 2022.

(要約): In natural indigo dyeing, the water-insoluble indigo included in the composted indigo leaves called sukumo is converted to water-soluble leuco-indigo through the reduction activities of microorganisms under alkaline conditions. To understand the relationship between indigo reduction and microorganisms in indigo-fermentation suspensions, we isolated and identified the microorganisms that reduce indigo and analyzed the microbiota in indigo-fermentation suspensions. Indigo-reducing microorganisms, which were not isolated by means of a conventional indigo carmine-reduction assay method, were isolated by using indigo as a direct substrate and further identified and characterized. We succeeded in isolating bacteria closely related to Corynebacterium glutamicum, Chryseomicrobium aureum, Enterococcus sp. for the first time. Anthraquinone was found to be an effective mediator that facilitated the indigo-reduction activity of the isolated strains. On analysis of the microbiota in indigo-fermentation suspensions, the ratio of indigo-reducing bacteria and others was found to be important for maintaining the indigo-reduction activity.
(キーワード): Indigo / Microbiota / 発酵 (fermentation)
(出版サイトへのリンク): ● Publication site (DOI): 10.1093/bbb/zbab209
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1093/bbb/zbab209

(DOI: 10.1093/bbb/zbab209) Naomi Murakawa, Takaiku Sakamoto, Mizuho Kanoh, Park Si-Bum, Shigenobu Kishino, Jun Ogawa and Eiji Sakuradani :
Microbial production of hydroxy fatty acids utilizing crude glycerol,
Biocatalysis and Agricultural Biotechnology, Vol.39, 102286, 2022.

(キーワード): Fusarium solani / 10-Hydroxyoctadecanoic acid / 10-Hydroxy-cis-12-octadecenoic acid / 10-Oxo-octadecanoic acid / Fatty acid hydration / Crude glycerol
(徳島大学機関リポジトリ): ● Metadata: 117466
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.bcab.2022.102286
(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1360013168859284992; ● Search Scopus @ Elsevier (DOI): 10.1016/j.bcab.2022.102286

(徳島大学機関リポジトリ: 117466, DOI: 10.1016/j.bcab.2022.102286, CiNii: 1360013168859284992) Takaiku Sakamoto, Yuichi Kamegawa, Chinami Kurita, Mizuho Kanoh, Naomi Murakawa and Eiji Sakuradani :
Efficient production of biolipids by crude glycerol-assimilating fungi,
Bioresource Technology Reports, Vol.16, 100861, 2021.

(要約): The aim of this study was to isolate microorganisms utilizing crude glycerol as a carbon source efficiently and to evaluate their lipid productivity. Fusarium oxysporum W1 grew well on medium containing 20% crude glycerol as well as 50% pure glycerol. The dry cell weights and total fatty acids of F. oxysporum W1 reached 24.5 g/L and 12.4 g/L. Penicillium sp. N1 and P. citrinum N3 were found to accumulate free fatty acids to as much as 56.2% and 48.5% of total fatty acids, respectively, on cultivation in the crude glycerol-containing medium. These strains grew well on medium containing crude glycerol only heat-treated at 80 105 °C without autoclave sterilization.
(キーワード): Crude glycerol / Biolipid / Penicillium / Fusarium
(徳島大学機関リポジトリ): ● Metadata: 117465
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.biteb.2021.100861
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1016/j.biteb.2021.100861

(徳島大学機関リポジトリ: 117465, DOI: 10.1016/j.biteb.2021.100861) Mayu Kikushi, Keisei Sowa, Kasumi Nakagawa, Momoka Matsunaga, Akinori Ando, Kenji Kano, Michiki Takeuchi and Eiji Sakuradani :
Indigo-mediated semi-microbial biofuel cell using an indigo-dye fermenting suspension,
Catalysts, Vol.11, No.9, 1080, 2021.

(要約): Aizome (Japanese indigo dyeing) is a unique dyeing method using microbial activity under anaerobic alkaline conditions. In indigo-dye fermenting suspensions; microorganisms reduce indigo into leuco-indigo with acetaldehyde as a reductant. In this study; we constructed a semi-microbial biofuel cell using an indigo-dye fermenting suspension. Carbon fiber and Pt mesh were used as the anode and cathode materials, respectively. The open-circuit voltage (OCV) was 0.6 V, and the maximum output power was 32 µW cm-2 (320 mW m-2). In addition, the continuous stability was evaluated under given conditions starting with the highest power density; the power density rapidly decreased in 0.5 h due to the degradation of the anode. Conversely, at the OCV, the anode potential exhibited high stability for two days. However, the OCV decreased by approximately 80 mV after 2 d compared with the initial value, which was attributed to the performance degradation of the gas-diffusion-cathode system caused by the evaporation of the dispersion solution. This is the first study to construct a semi-microbial biofuel cell using an indigo-dye fermenting suspension.
(キーワード): indigo / indigo fermentation / microbial fuel cell
(徳島大学機関リポジトリ): ● Metadata: 116722
(出版サイトへのリンク): ● Publication site (DOI): 10.3390/catal11091080
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.3390/catal11091080

(徳島大学機関リポジトリ: 116722, DOI: 10.3390/catal11091080) Kasumi Nakagawa, Michiki Takeuchi, Mayu Kikuchi, Manami Tada, Takaiku Sakamoto, Kenji Kano, Jun Ogawa and Eiji Sakuradani :
Voltammetric in-situ monitoring of leuco-indigo in indigo-fermenting suspensions,
Journal of Bioscience and Bioengineering, Vol.131, No.5, 565-571, 2021.

(要約): Cyclic voltammetry was successfully applied to in-vivo monitoring of leuco-indigo in indigo-fermenting suspensions under quiescent conditions without deoxygenation; the working and counter electrodes were kept on the surface of each suspension by a polyethylene vinyl alcohol tube holder. The anodic peak current was used as a measure of the leuco-indigo concentration. The voltammetric wave shape suggested partial solubilization of the indigo with some macromolecules in the fermenting suspensions, which lead to an in-situ method without any electrode surface pretreatment. The anodic peak current well reflected the dyeing activity of a suspensions. The results obtained for laboratory-level fermentation systems clarified the number of days required for dye fermentation, the effectiveness of addition of old suspension as an additive for preparing fresh fermenting suspensions, and the importance of addition of a nitrogen-based nutrient as well as a glucose-based one to recover the indigo-reducing activity. The method can also be applied to determine the amounts of indigo in used dye suspensions and extracts of fermented indigo leaves (sukumo) by adding a chemical reduction pretreatment.
(キーワード): Indigo / 発酵 (fermentation) / 電気化学 (electrochemistry) / Anaerobic condition / Cyclic voltammetry
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2021.01.005
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2021.01.005

(DOI: 10.1016/j.jbiosc.2021.01.005) Brian KH Mo, Akinori Ando, Ryohei Nakatsuji, Tomoyo Okuda, Yuki Takemoto, Hiroyuki Ikemoto, Hiroshi Kikukawa, Takaiku Sakamoto, Eiji Sakuradani and Jun Ogawa :
Characterization of ω3 fatty acid desaturases from oomycetes and their application toward eicosapentaenoic acid production in Mortierella alpina,
Bioscience, Biotechnology, and Biochemistry, Vol.85, No.5, 1252-1265, 2021.

(要約): ω3 polyunsaturated fatty acids are currently obtained mainly from fisheries, thus sustainable alternative sources such as oleaginous microorganisms are required. Here we describe the isolation, characterization, and application of three novel ω3 desaturases with ω3 polyunsaturated fatty acid-producing activity at ordinary temperatures (28 °C). First, we selected Pythium sulcatum and Plectospira myriandra after screening for oomycetes with high eicosapentaenoic acid/arachidonic acid ratios and isolated the genes psulω3 and pmd17, respectively, which encode ω3 desaturases. Subsequent characterization showed that PSULω3 exhibited ω3 desaturase activity on both C18 and C20 ω6 polyunsaturated fatty acids while PMD17 exhibited ω3 desaturase activity exclusively on C20 ω6 polyunsaturated fatty acids. Expression of psulω3 and pmd17 in the arachidonic acid-producer Mortierella alpina resulted in transformants that produced eicosapentaenoic acid/total fatty acid values of 38% and 40%, respectively, at ordinary temperatures. These ω3 desaturases should facilitate the construction of sustainable ω3 polyunsaturated fatty acid sources.
(キーワード): Eicosapentaenoic acid / Pythium sulcatum / Plectospira myriandra / ω3 desaturase / Mortierella alpina
(出版サイトへのリンク): ● Publication site (DOI): 10.1093/bbb/zbaa123
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1093/bbb/zbaa123

(DOI: 10.1093/bbb/zbaa123) Kasumi Nakagawa, Michiki Takeuchi, Mayu Kikuchi, Suzuna Kiyofuji, Masami Kugo, Takaiku Sakamoto, Kenji Kano, Jun Ogawa and Eiji Sakuradani :
Mechanistic Insights into Indigo Reduction in Indigo Fermentation: A Voltammetric Study,
Electrochemistry, Vol.89, 25-30, 2021.

(要約): Indigo is one of the oldest natural blue dyes. Microorganisms and their enzymatic activities are deeply involved in the traditional indigo staining procedure. To elucidate the mechanism of the microbial indigo reduction, we directly performed cyclic voltammetry on alkaline fermenting dye suspensions. A pair of characteristic redox peaks of leuco-indigo was observed in a supernatant fluid of the fermenting dye suspension. On the other hand, it was found that the indigo/leuco-indigo redox couple mediated two-way microbially catalyzed oxidation and reduction in a sediment-rich suspension of the fermenting suspension. Acetaldehyde was supposed to be the electron donor and acceptor of the catalytic reactions. In order to verify the bioelectrocatalytic reaction, we isolated indigo-reducing bacterium K2-3 from the fermenting suspension, and the two-way bioelectrocatalysis was successfully restaged in a model system containing K2-3 and methyl viologen (as a soluble mediator instead of indigo) as well as acetaldehyde at pH 10.
(キーワード): Indigo Reduction Mechanism / Acetaldehyde / Bioelectrocatalysis / Mediator
(徳島大学機関リポジトリ): ● Metadata: 116295
(出版サイトへのリンク): ● Publication site (DOI): 10.5796/electrochemistry.20-00123
(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1390568456342112896; ● Summary page in Scopus @ Elsevier: 2-s2.0-85099770189

(徳島大学機関リポジトリ: 116295, DOI: 10.5796/electrochemistry.20-00123, CiNii: 1390568456342112896, Elsevier: Scopus) Takeru Koga, Takaiku Sakamoto, Eiji Sakuradani and Akihiro Tai :
Neurite Outgrowth-Promoting Activity of Compounds in PC12 Cells from Sunflower Seeds,
Molecules, Vol.25, No.20, 4748, 2020.

(要約): In the current super-aging society, the establishment of methods for prevention and treatment of Alzheimer's disease (AD) is an urgent task. One of the causes of AD is thought to be a decrease in the revel of nerve growth factor (NGF) in the brain. Compounds showing NGF-mimicking activity and NGF-enhancing activity have been examined as possible agents for improving symptoms. In the present study, sunflower seed extract was found to have neurite outgrowth-promoting activity, which is an NGF-enhancing activity, in PC12 cells. To investigate neurite outgrowth-promoting compounds from sunflower seed extract, bioassay-guided purification was carried out. The purified active fraction was obtained by liquid-liquid partition followed by some column chromatographies. Proton nuclear magnetic resonance and gas chromatography-mass spectrometry analyses of the purified active fraction indicated that the fraction was a mixture of β-sitosterol, stigmasterol and campesterol, with β-sitosterol being the main component. Neurite outgrowth-promoting activities of β-sitosterol, stigmasterol, campesterol and cholesterol were evaluated in PC12 cells. β-Sitosterol and stigmasterol showed the strongest activity of the four sterol compounds (β-sitosterol stigmasterol > campesterol > cholesterol), and cholesterol did not show any activity. The results indicated that β-sitosterol was the major component responsible for the neurite outgrowth-promoting activity of sunflower seeds. Results of immunostaining also showed that promotion by β-sitosterol of neurite formation induced by NGF was accompanied by neurofilament expression. β-Sitosterol, which showed NGF-enhancing activity, might be a candidate ingredient in food for prevention of AD.
(キーワード): Alzheimer's disease / sunflower seed / NGF / β-sitosterol / neurite outgrowth-promoting activit
(徳島大学機関リポジトリ): ● Metadata: 116242
(出版サイトへのリンク): ● Publication site (DOI): 10.3390/molecules25204748
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.3390/molecules25204748

(徳島大学機関リポジトリ: 116242, DOI: 10.3390/molecules25204748) Takaiku Sakamoto, Eiji Sakuradani, Tomoyo Okuda, Hiroshi Kikukawa, Akinori Ando, Shigenobu Kishino, Yoshihiro Izumi, Takeshi Bamba, Jun Shima and Jun Ogawa :
Metabolic engineering of oleaginous fungus Mortierella alpina for high production of oleic and linoleic acids,
Bioresource Technology, Vol.245, 1610-1615, 2017.

(要約): The aim of this work was to study the molecular breeding of oleaginous filamentous Mortierella alpina for high production of linoleic (LA) or oleic acid (OA). Heterologous expression of the Δ12-desaturase (DS) gene derived from Coprinopsis cinerea in the Δ6DS activity-defective mutant of M. alpina increased the LA production rate as to total fatty acid to 5 times that in the wild strain. By suppressing the endogenous Δ6I gene expression by RNAi in the Δ12DS activity-defective mutant of M. alpina, the OA accumulation rate as to total fatty acid reached 68.0%. The production of LA and OA in these transformants reached 1.44 and 2.76g/L, respectively, on the 5th day. The Δ6I transcriptional levels of the RNAi-treated strains were suppressed to 1/10th that in the parent strain. The amount of Δ6II RNA in the Δ6I RNAi-treated strain increased to 8 times that in the wild strain.
(徳島大学機関リポジトリ): ● Metadata: 113896
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.biortech.2017.06.089
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 28673516; ● Search Scopus @ Elsevier (PMID): 28673516; ● Search Scopus @ Elsevier (DOI): 10.1016/j.biortech.2017.06.089

(徳島大学機関リポジトリ: 113896, DOI: 10.1016/j.biortech.2017.06.089, PubMed: 28673516) Hiroshi Kikukawa, Eiji Sakuradani, Akinori Ando, Tomoyo Okuda, Sakayu Shimizu and Jun Ogawa :
Microbial production of dihomo-γ-linolenic acid by Δ5-desaturase gene-disruptants of Mortierella alpina 1S-4.,
Journal of Bioscience and Bioengineering, Vol.122, No.1, 22-26, 2016.

(要約): We constructed dihomo-γ-linolenic acid (DGLA)-producing strains with disruption of the Δ5-desaturase (Δ5ds) gene, which encodes a key enzyme catalyzing the bioconversion of DGLA to arachidonic acid (ARA), by efficient gene-targeting, using Δlig4 strain of Mortierella alpina 1S-4 as the host. In previous study, we had already identified and disrupted the lig4 gene encoding DNA ligase 4, which involves in non-homologous end joining, in M. alpina 1S-4, and the Δlig4 strain had showed efficient gene-targeting. In this study, the uracil auxotroph of Δlig4 strain was constructed, and then transformed for disruption of Δ5ds. The isolation of nine Δ5ds-disruptants out of 18 isolates indicated that the disruption efficiency was 50%. The ratio of DGLA among the total fatty acids of the Δ5ds-disruptants reached 40.1%; however, no ARA was detected. To our knowledge, this is the first study to report the construction of DGLA-producing transformants by using the efficient gene-targeting system in M. alpina 1S-4.
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2015.12.007
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 26777235; ● Search Scopus @ Elsevier (PMID): 26777235; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2015.12.007

(DOI: 10.1016/j.jbiosc.2015.12.007, PubMed: 26777235) Tomohiro Fujita, Hiroshi Kawashima, Eiji Sakuradani, Takaiku Sakamoto, Akinori Ando, Jun Ogawa and Sakayu Shimizu :
Essential fatty acids for oleaginous fungus Mortierella alpina,
Biocatalysis and Agricultural Biotechnology, Vol.8, 167-170, 2016.

(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.bcab.2016.08.011
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1016/j.bcab.2016.08.011

(DOI: 10.1016/j.bcab.2016.08.011) Tomoyo Okuda, Akinori Ando, Hiroaki Negoro, Tatsuya Muratsubaki, Hiroshi Kikukawa, Takaiku Sakamoto, Eiji Sakuradani, Sakayu Shimizu and Jun Ogawa :
Eicosapentaenoic acid (EPA) production by an oleaginous fungus Mortierella alpina expressing heterologous the Δ17 desaturase gene under ordinary temperature,
European Journal of Lipid Science and Technology : EJLST, Vol.117, No.12, 1919-1927, 2015.

(要約): The oleaginous fungus Mortierella alpina is known to accumulate eicosapentaenoic acid (EPA) only when cultivated at a low temperature (below 15°C). Here, we investigated EPA production at a ordinary temperature (28°C) by expressing the Saprolegnia diclina 17 desaturase gene (sdd17m) in M. alpina ST1358, an 3-desaturation activity-defective mutant derived from M. alpina 1S-4. Expression of the exogenous gene was confirmed by EPA accumulation in transformants at both 28°C and 12°C. The EPA content in total lipids produced by transformants was over 20% at 28°C. Bench-scale fermentation with a 5-L jar fermentor showed that EPA content reached 26.4% of total fatty acids, and final EPA production reached 1.8 g/L. This is the first study to report the accumulation of EPA in M. alpina at a ordinary temperature, and provide a platform technology for the industrial production of EPA using M. alpina as a promising source for EPA.
(キーワード): 3 polyunsaturated fatty acid / ordinary temperature / Mortierella alpina / Saprolegnia diclina
(出版サイトへのリンク): ● Publication site (DOI): 10.1002/ejlt.201400657
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1002/ejlt.201400657

(DOI: 10.1002/ejlt.201400657) Hiroshi Kikukawa, Eiji Sakuradani, Akinori Ando, Tomoyo Okuda, Misa Ochiai, Sakayu Shimizu and Jun Ogawa :
Disruption of lig4 improves gene targeting efficiency in the oleaginous fungus Mortierella alpina 1S-4.,
Journal of Biotechnology, Vol.208, 63-69, 2015.

(要約): The oil-producing zygomycete Mortierella alpina 1S-4 is known to accumulate beneficial polyunsaturated fatty acids. We identified the lig4 gene that encodes for a DNA ligase 4 homolog, which functions to repair double strand breaks by non-homologous end joining. We disrupted the lig4 gene to improve the gene targeting efficiency in M. alpina. The M. alpina 1S-4 Δlig4 strains showed no defect in vegetative growth, formation of spores, and fatty acid production, but exhibited high sensitivity to methyl methansulfonate, an agent that causes DNA double-strand breaks. Importantly, gene replacement of ura5 marker by CBXB marker occurred in 67% of Δlig4 strains and the gene targeting efficiency was 21-fold greater than that observed in disruption of the lig4 gene in the M. alpina 1S-4 host strain. Further metabolic engineering of the Δlig4 strains is expected to result in strains that produce higher levels of rare and beneficial polyunsaturated fatty acids and contribute to basic research on the zygomycete.
(キーワード): DNA Ligases / Fungal Proteins / Gene Knockdown Techniques / Gene Targeting / Mortierella
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiotec.2015.05.020
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 26052021; ● Summary page in Scopus @ Elsevier: 2-s2.0-84930934381

(DOI: 10.1016/j.jbiotec.2015.05.020, PubMed: 26052021, Elsevier: Scopus) Tomoyo Okuda, Akinori Ando, Hiroaki Negoro, Hiroshi Kikukawa, Takaiku Sakamoto, Eiji Sakuradani, Sakayu Shimizu and Jun Ogawa :
Omega-3 eicosatetraenoic acid production by molecular breeding of the mutant strain S14 derived from Mortierella alpina 1S-4.,
Journal of Bioscience and Bioengineering, Vol.120, No.3, 299-304, 2015.

(要約): We investigated the omega-3 eicosatetraenoic acid (ETA) production by molecular breeding of the oleaginous fungus Mortierella alpina, which can slightly accumulate ETA only when cultivated at a low temperature. The endogenous ω3-desaturase gene or the heterologous Saprolegnia diclina Δ17 (sdd17m) desaturase gene were overexpressed in M. alpina S14, a Δ5-desaturation activity-defective mutant derived from M. alpina 1S-4. M. alpina S14 transformants introduced with the endogenous ω3-desaturase gene showed ETA at 42.1% content in the total lipids that was 84.2-fold and 3.2-fold higher than that of the wild-type strain 1S-4 and host strain S14, respectively, when cultivated at 12°C. No accumulation of ETA was observed at 28°C. In contrast, transformants with the heterologous sdd17m gene showed 24.9% of the content of total lipids at 28°C. These results indicated that these M. alpina S14 transformants are promising strains for the production of ETA, which is hard to obtain from natural sources.
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2015.01.014
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 25845716; ● Summary page in Scopus @ Elsevier: 2-s2.0-84938292293

(DOI: 10.1016/j.jbiosc.2015.01.014, PubMed: 25845716, Elsevier: Scopus) Hiroshi Kikukawa, Eiji Sakuradani, Masato Nakatani, Akinori Ando, Tomoyo Okuda, Takaiku Sakamoto, Misa Ochiai, Sakayu Shimizu and Jun Ogawa :
Gene targeting in the oil-producing fungus Mortierella alpina 1S-4 and construction of a strain producing a valuable polyunsaturated fatty acid.,
Current Genetics, Vol.61, No.4, 579-589, 2015.

(要約): To develop an efficient gene-targeting system in Mortierella alpina 1S-4, we identified the ku80 gene encoding the Ku80 protein, which is involved in the nonhomologous end-joining pathway in genomic double-strand break (DSB) repair, and constructed ku80 gene-disrupted strains via single-crossover homologous recombination. The ku80 strain from M. alpina 1S-4 showed no negative effects on vegetative growth, formation of spores, and fatty acid productivity, and exhibited high sensitivity to methyl methanesulfonate, which causes DSBs. Dihomo--linolenic acid (DGLA)-producing strains were constructed by disruption of the 5-desaturase gene, encoding a key enzyme of bioconversion of DGLA to ARA, using the ku80 strain as a host strain. The significant improvement of gene-targeting efficiency was not observed by disruption of the ku80 gene, but the construction of DGLA-producing strain by disruption of the 5-desaturase gene was succeeded using the ku80 strain as a host strain. This report describes the first study on the identification and disruption of the ku80 gene in zygomycetes and construction of a DGLA-producing transformant using a gene-targeting system in M. alpina 1S-4.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00294-015-0481-2
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 25782448; ● Summary page in Scopus @ Elsevier: 2-s2.0-84944513241

(DOI: 10.1007/s00294-015-0481-2, PubMed: 25782448, Elsevier: Scopus) H Kikukawa, Eiji Sakuradani, Y Nishibaba, T Okuda, A Ando, J Shima, S Shimizu and J Ogawa :
Production of cis-11-eicosenoic acid by Mortierella fungi.,
Journal of Applied Microbiology, Vol.118, No.3, 641-647, 2015.

(要約): We found that M. chlamydospora CBS 529.75 was the best producer for the microbial production of EA.
(出版サイトへのリンク): ● Publication site (DOI): 10.1111/jam.12725
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 25495454; ● Summary page in Scopus @ Elsevier: 2-s2.0-84926156274

(DOI: 10.1111/jam.12725, PubMed: 25495454, Elsevier: Scopus) Eiji Sakuradani, Hiroshi Kikukawa, Seiki Takeno, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Transformation of zygomycete Mortierella alpina using biolistic particle bombardment,
Genetic Transformation Systems in Fungi., Vol.1, 135-140, 2014.

(出版サイトへのリンク): ● Publication site (DOI): 10.1007/978-3-319-10142-2_13
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1007/978-3-319-10142-2_13

(DOI: 10.1007/978-3-319-10142-2_13) Ayumi Tanimura, Masako Takashima, Takashi Sugita, Rikiya Endoh, Minako Kikukawa, Shino Yamaguchi, Eiji Sakuradani, Jun Ogawa, Moriya Ohkuma and Jun Shima :
Cryptococcus terricola is a promising oleaginous yeast for biodiesel production from starch through consolidated bioprocessing.,
Scientific Reports, Vol.4, 4776, 2014.

(要約): Starch is considered a potential feedstock for biofuel production, particularly in light of the large-scale landfilling of food waste and other starchy materials worldwide. Lipid accumulation by oleaginous yeast is a promising method for biodiesel production from starch. However, most oleaginous yeasts are grown on monosaccharides or oligosaccharides because they cannot directly utilize starch. We therefore investigated the starch-assimilation ability of 1,200 yeasts. We found that Cryptococcus terricola could be used for fuel production through consolidated bioprocessing. C. terricola JCM 24523 exhibited the highest lipid content of 61.96% on medium with 5% starch at 10 days. Fatty acid methyl ester analysis showed that this strain produced high proportions of C16:0 and C18 fatty acids when grown on starch, which are ideal for use in biodiesel. Considering the yield and cost, lipids derived from starch using C. terricola would be a promising alternative source for biodiesel production.
(キーワード): Biofuels / Cryptococcus / Culture Media / Environmental Microbiology / Fatty Acids / Industrial Microbiology / Kinetics / Lipid Metabolism / Starch
(出版サイトへのリンク): ● Publication site (DOI): 10.1038/srep04776
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24758986; ● Search Scopus @ Elsevier (PMID): 24758986; ● Search Scopus @ Elsevier (DOI): 10.1038/srep04776

(DOI: 10.1038/srep04776, PubMed: 24758986) Tomoyo Okuda, Akinori Ando, Eiji Sakuradani, Hiroshi Kikukawa, Nozomu Kamada, Misa Ochiai, Jun Shima and Jun Ogawa :
Selection and characterization of promoters based on genomic approach for the molecular breeding of oleaginous fungus Mortierella alpina 1S-4.,
Current Genetics, Vol.60, No.3, 183-191, 2014.

(要約): To express a foreign gene effectively, a good expression system is required. In this study, we investigated various promoters as useful tools for gene manipulation in oleaginous fungus Mortierella alpina 1S-4. We selected and cloned the promoter regions of 28 genes in M. alpina 1S-4 on the basis of expression sequence tag abundance data. The activity of each promoter was evaluated using the -glucuronidase (GUS) reporter gene. Eight of these promoters were shown to enhance GUS expression more efficiently than a histone promoter, which is conventionally used for the gene manipulation in M. alpina. Especially, the predicted protein 3 and the predicted protein 6 promoters demonstrated approximately fivefold higher activity than the histone promoter. The activity of some promoters changed along with the cultivation phase of M. alpina 1S-4. Seven promoters with constitutive or time-dependent, high-level expression activity were selected, and deletion analysis was carried out to determine the promoter regions required to retain activity. This is the first report of comprehensive promoter analysis based on a genomic approach for M. alpina. The promoters described here will be useful tools for gene manipulation in this strain.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00294-014-0423-4
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24562865; ● Search Scopus @ Elsevier (PMID): 24562865; ● Search Scopus @ Elsevier (DOI): 10.1007/s00294-014-0423-4

(DOI: 10.1007/s00294-014-0423-4, PubMed: 24562865) Tomoyo Okuda, Akinori Ando, Eiji Sakuradani, Hiroshi Kikukawa, Nozomu Kamada, Misa Ochiai, Jun Shima and Jun Ogawa :
Characterization of galactose-dependent promoters from an oleaginous fungus Mortierella alpina 1S-4.,
Current Genetics, Vol.60, No.3, 175-182, 2014.

(要約): An inducible promoter is a useful tool for the controlled expression of a given gene. In this report, we describe galactose-dependent promoters for potential use in an oleaginous fungus Mortierella alpina. We cloned the putative promoter regions of two genes encoding galactose metabolic enzymes, GAL1 and GAL10, from the genome of M. alpina 1S-4. The -glucuronidase (GUS) reporter gene assay in M. alpina 1S-4 revealed that regulation of these promoters was dependent on the presence of galactose in the medium both with and without other sugars. With the GAL10 promoter, an approximately 50-fold increase of GUS activity was demonstrated by addition of galactose into the culture media at any cultivation phase. The 5' deletion analysis of the GAL10 promoter revealed that a promoter region of over 2,000 bp length was required for its high-level activity and sufficient inducible response. Significantly, this is the first report of inducible promoters of zygomycetes. The GAL10 promoter will be a valuable tool for gene manipulation in M. alpina 1S-4.
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00294-014-0422-5
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24549368; ● Search Scopus @ Elsevier (PMID): 24549368; ● Search Scopus @ Elsevier (DOI): 10.1007/s00294-014-0422-5

(DOI: 10.1007/s00294-014-0422-5, PubMed: 24549368) Ayumi Tanimura, Masako Takashima, Takashi Sugita, Rikiya Endoh, Minako Kikukawa, Shino Yamaguchi, Eiji Sakuradani, Jun Ogawa and Jun Shima :
Selection of oleaginous yeasts with high lipid productivity for practical biodiesel production.,
Bioresource Technology, Vol.153, 230-235, 2013.

(要約): The lipid-accumulating ability of 500 yeast strains isolated in Japan was evaluated. Primary screening revealed that 31 strains were identified as potential lipid producers, from which 12 strains were cultivated in a medium containing 3% glucose. It was found that JCM 24511 accumulated the highest lipid content, up to 61.53%, while JCM 24512 grew the fastest. They were tentatively identified as Cryptococcus sp. and Cryptococcus musci, respectively. The maximum lipid concentration of 1.49g/L was achieved by JCM 24512. Similarly, JCM 24511 also achieved a high lipid production of 1.37g/L. High lipid productivity is the most important characteristic of oleaginous yeasts from the viewpoint of practical production. Among the strains tested here, JCM 24512 had the best lipid productivity, 0.37g/L/day. The results show that the isolated yeasts could be promising candidates for biodiesel production.
(キーワード): Biofuels / Biomass / Biotechnology / Cryptococcus / European Union / Fatty Acids / Jatropha / Kinetics / Lipids / Plant Oils / Time Factors / United States
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.biortech.2013.11.086
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24368271; ● Search Scopus @ Elsevier (PMID): 24368271; ● Search Scopus @ Elsevier (DOI): 10.1016/j.biortech.2013.11.086

(DOI: 10.1016/j.biortech.2013.11.086, PubMed: 24368271) Yasushi Takimura, Eiji Sakuradani, Yusuke Natsume, Takashi Miyake, Jun Ogawa and Sakayu Shimizu :
Achlorophyllous alga Prototheca zopfii oxidizes n-alkanes with different carbon-chain lengths through a unique subterminal oxidation pathway.,
Journal of Bioscience and Bioengineering, Vol.117, No.3, 275-277, 2013.

(要約): Some Prototheca spp. were previously reported to convert n-hexadecane to 5-hexadecanol and then to 5-hexadecanone through a unique subterminal oxidation pathway. Further analysis of derivatives derived from n-hexadecane indicated that Prototheca zopfii oxidized n-alkanes with C11 to C17 chain lengths at not only the 5th but also the 4th, 3rd and 2nd positions.
(キーワード): Alkanes / Carbon / Gas Chromatography-Mass Spectrometry / Oxidation-Reduction / Prototheca
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2013.09.004
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 24099955; ● Search Scopus @ Elsevier (PMID): 24099955; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2013.09.004

(DOI: 10.1016/j.jbiosc.2013.09.004, PubMed: 24099955) Hiroshi Kikukawa, Eiji Sakuradani, Shigenobu Kishino, Si-Bum Park, Akinori Ando, Jun Shima, Misa Ochiai, Sakayu Shimizu and Jun Ogawa :
Characterization of a trifunctional fatty acid desaturase from oleaginous filamentous fungus Mortierella alpina 1S-4 using a yeast expression system.,
Journal of Bioscience and Bioengineering, Vol.116, No.6, 672-676, 2013.

(要約): A ω3-fatty acid desaturase gene (maw3) which is involved in biosynthesis of n-3 polyunsaturated fatty acids (PUFAs) was previously isolated from Mortierella alpina 1S-4. In this report, we investigated the products of MAW3 catalyzing reaction with endogenous and exogenous fatty acids in the yeast transformant. Two unusual fatty acids de novo synthesized in the yeast transformant expressing maw3 gene were identified as n-4 hexadecadienoic acid (16:2(9cis,12cis)) and n-1 hexadecatrienoic acid (16:3(9cis,12cis,15)) by GC-MS and (1)H NMR analyses. In addition to the desaturation activity at the ω3-position for 18- and 20-carbon PUFAs, MAW3 in the yeast transformant inserted a double bond at Δ12-position of endogenous palmitoleic acid (16:1(9cis)) and further at Δ15-position of the resulting 16:2(9cis,12cis) to result in the formation of 16:3(9cis,12cis,15) leading to a bifunctional Δ12/Δ15-desaturase for 16-carbon fatty acids. Moreover, we evaluated the activity of MAW3 in the yeast transformant under different temperatures. The MAW3 did not have desaturation activities in M. alpina 1S-4 at 28°C but it had in the yeast transformant for various fatty acids. The MAW3 was demonstrated to be a trifunctional Δ12/Δ15/ω3-desaturase, exhibiting Δ12-desaturation for 16:1(9cis), Δ15-desaturation for 16- and 18-carbon fatty acids that had a preexisting cis-double bond at Δ12 position, and ω3-desaturation for 20-carbon fatty acids having that at Δ14-position. It is the first report that the fatty acid desaturase (MAW3) is shown to have Δ12- and Δ15-desaturation activities for a 16-carbon fatty acid, in addition to its major function, ω3-desaturation activity.
(キーワード): Fatty Acid Desaturases / Fatty Acids, Monounsaturated / Fatty Acids, Unsaturated / 遺伝子発現 (gene expression) / Mortierella / Nuclear Magnetic Resonance, Biomolecular / Saccharomyces cerevisiae
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2013.05.023
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23790547; ● Search Scopus @ Elsevier (PMID): 23790547; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2013.05.023

(DOI: 10.1016/j.jbiosc.2013.05.023, PubMed: 23790547) Eiji Sakuradani, Yusuke Natsume, Yasushi Takimura, Jun Ogawa and Sakayu Shimizu :
Subterminal oxidation of n-alkanes in achlorophyllous alga Prototheca sp.,
Journal of Bioscience and Bioengineering, Vol.116, No.4, 472-474, 2013.

(要約): Some Prototheca sp. are known to be involved in n-hexadecane degradation. Two derivatives derived from n-hexadecane in such Prototheca sp. were identified as 5-hexadecanone and 5-hexadecanol. n-Hexadecane was assumed to be converted to 5-hexadecanol and then to 5-hexadecanone through a unique subterminal oxidation pathway in such Prototheca sp.
(キーワード): Alkanes / Fatty Alcohols / Oxidation-Reduction / Prototheca
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2013.04.019
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23651808; ● Search Scopus @ Elsevier (PMID): 23651808; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2013.04.019

(DOI: 10.1016/j.jbiosc.2013.04.019, PubMed: 23651808) Eiji Sakuradani, Lifang Zhao, M Tegan Haslam and Ljerka Kunst :
The CER22 gene required for the synthesis of cuticular wax alkanes in Arabidopsis thaliana is allelic to CER1.,
Planta, Vol.237, No.3, 731-738, 2012.

(要約): Cuticular waxes coat the primary aerial tissues of land plants and serve as a protective barrier against non-stomatal water loss and various environmental stresses. Alkanes are the most prominent cuticular wax components and are thought to have an important role in controlling permeability of the cuticle. However, alkane biosynthesis in plants is not well understood. Arabidopsis eceriferum1 (cer1) and cer22 mutants show dramatic reductions in alkane, secondary alcohol, and ketone content, and concomitant increases in aldehyde content, suggesting that one or both of these genes encode an alkane-forming enzyme. To determine the biochemical identity of CER22, and to investigate the relationship between CER1 and CER22 in alkane formation, we mapped the cer22 mutation as a first step to positional cloning. Unexpectedly, mapping revealed linkage of cer22 to markers on chromosome 1 in the vicinity of CER1, and not to markers on chromosome 3 as previously reported. Failure of the cer1-1 and cer22 mutants to complement each other, and the presence of an allele specific mutation in the CER1 gene amplified from cer22 genomic DNA demonstrated that CER22 is identical to CER1. The cer22 mutant was therefore renamed cer1-6. Analyses of CER1 transcript levels, and stem cuticular wax load and composition in the cer1-6 (cer22) line indicated that cer1-6 is a weak mutant allele of CER1. This represents an important step forward in our understanding of alkane synthesis in plants, and will direct future research in the field to focus on the role of CER1 in this process.
(キーワード): Alkanes / Alleles / Arabidopsis / Arabidopsis Proteins / Chromosomes, Plant / Gene Expression Regulation, Plant / Genes, Plant / Genetic Complementation Test / Mutation / Physical Chromosome Mapping / Plant Epidermis / Plant Stems / Waxes
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00425-012-1791-y
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23117394; ● Search Scopus @ Elsevier (PMID): 23117394; ● Search Scopus @ Elsevier (DOI): 10.1007/s00425-012-1791-y

(DOI: 10.1007/s00425-012-1791-y, PubMed: 23117394) Chikara Ohto, Masayoshi Muramatsu, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Production of geranylgeraniol on overexpression of a prenyl diphosphate synthase fusion gene in Saccharomyces cerevisiae.,
Applied Microbiology and Biotechnology, Vol.87, No.4, 1327-1334, 2010.

(要約): An acyclic diterpene alcohol, (E,E,E)-geranylgeraniol (GGOH), is one of the important compounds used as perfume and pharmacological agents. A deficiency of squalene (SQ) synthase activity allows yeasts to accumulate an acyclic sesquiterpene alcohol, (E,E)-farnesol, in their cells. Since sterols are essential for the growth of yeasts, a deficiency of SQ synthase activity makes the addition of supplemental sterols to the culture media necessary. To develop a GGOH production method not requiring any supplemental sterols, we overexpressed HMG1 encoding hydroxymethylglutaryl-CoA reductase and the genes of two prenyl diphosphate synthases, ERG20 and BTS1, in Saccharomyces cerevisiae. A prototrophic diploid coexpressing HMG1 and the ERG20-BTS1 fusion accumulated GGOH with neither disruption of the SQ synthase gene nor the addition of any supplemental sterols. The GGOH content on the diploid cultivation in a 5-l jar fermenter reached 138.8 mg/l under optimal conditions.
(キーワード): Diterpenes / Gene Expression / Geranyltranstransferase / Hydroxymethylglutaryl CoA Reductases / Recombinant Fusion Proteins / Saccharomyces cerevisiae / Saccharomyces cerevisiae Proteins
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-010-2571-x
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 20393702; ● Search Scopus @ Elsevier (PMID): 20393702; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-010-2571-x

(DOI: 10.1007/s00253-010-2571-x, PubMed: 20393702) Nobuto Matsushita, Masahiro Miyashita, Yayoi Ichiki, Takehiko Ogura, Eiji Sakuradani, Yoshiaki Nakagawa, Sakayu Shimizu and Hisashi Miyagawa :
Purification and cDNA cloning of LaIT2, a novel insecticidal toxin from venom of the scorpion Liocheles australasiae.,
Bioscience, Biotechnology, and Biochemistry, Vol.73, No.12, 2769-2772, 2009.

(要約): The novel insecticidal toxin, LaIT2, was isolated from venom of the scorpion Liocheles australasiae. The amino acid sequence of LaIT2 was determined by an Edman degradation analysis and subsequent cDNA cloning. LaIT2 is composed of 59 amino acids with three disulfide bridges, and shares sequence similarity to the scorpion beta-KTx peptides.
(キーワード): Amino Acid Sequence / Animals / Base Sequence / Cloning, Molecular / DNA, Complementary / Insecticides / Molecular Sequence Data / Scorpion Venoms / Scorpions
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.90509
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19966481; ● Search Scopus @ Elsevier (PMID): 19966481; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.90509

(DOI: 10.1271/bbb.90509, PubMed: 19966481) Kenro Tokuhiro, Masayoshi Muramatsu, Chikara Ohto, Toshiya Kawaguchi, Shusei Obata, Nobuhiko Muramoto, Masana Hirai, Haruo Takahashi, Akihiko Kondo, Eiji Sakuradani and Sakayu Shimizu :
Overproduction of geranylgeraniol by metabolically engineered Saccharomyces cerevisiae.,
Applied and Environmental Microbiology, Vol.75, No.17, 5536-5543, 2009.

(要約): (E, E, E)-Geranylgeraniol (GGOH) is a valuable starting material for perfumes and pharmaceutical products. In the yeast Saccharomyces cerevisiae, GGOH is synthesized from the end products of the mevalonate pathway through the sequential reactions of farnesyl diphosphate synthetase (encoded by the ERG20 gene), geranylgeranyl diphosphate synthase (the BTS1 gene), and some endogenous phosphatases. We demonstrated that overexpression of the diacylglycerol diphosphate phosphatase (DPP1) gene could promote GGOH production. We also found that overexpression of a BTS1-DPP1 fusion gene was more efficient for producing GGOH than coexpression of these genes separately. Overexpression of the hydroxymethylglutaryl-coenzyme A reductase (HMG1) gene, which encodes the major rate-limiting enzyme of the mevalonate pathway, resulted in overproduction of squalene (191.9 mg liter(-1)) rather than GGOH (0.2 mg liter(-1)) in test tube cultures. Coexpression of the BTS1-DPP1 fusion gene along with the HMG1 gene partially redirected the metabolic flux from squalene to GGOH. Additional expression of a BTS1-ERG20 fusion gene resulted in an almost complete shift of the flux to GGOH production (228.8 mg liter(-1) GGOH and 6.5 mg liter(-1) squalene). Finally, we constructed a diploid prototrophic strain coexpressing the HMG1, BTS1-DPP1, and BTS1-ERG20 genes from multicopy integration vectors. This strain attained 3.31 g liter(-1) GGOH production in a 10-liter jar fermentor with gradual feeding of a mixed glucose and ethanol solution. The use of bifunctional fusion genes such as the BTS1-DPP1 and ERG20-BTS1 genes that code sequential enzymes in the metabolic pathway was an effective method for metabolic engineering.
(キーワード): Biosynthetic Pathways / Diterpenes / Farnesyltranstransferase / Genetic Engineering / Geranyltranstransferase / Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent / Membrane Proteins / Models, Biological / Pyrophosphatases / Recombinant Fusion Proteins / Saccharomyces cerevisiae / Saccharomyces cerevisiae Proteins / Squalene
(出版サイトへのリンク): ● Publication site (DOI): 10.1128/AEM.00277-09
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19592534; ● Search Scopus @ Elsevier (PMID): 19592534; ● Search Scopus @ Elsevier (DOI): 10.1128/AEM.00277-09

(DOI: 10.1128/AEM.00277-09, PubMed: 19592534) Akinori Ando, Yosuke Sumida, Hiroaki Negoro, Anggraini Dian Suroto, Jun Ogawa, Eiji Sakuradani and Sakayu Shimizu :
Establishment of Agrobacterium tumefaciens-mediated transformation of an oleaginous fungus, Mortierella alpina 1S-4, and its application for eicosapentaenoic acid producer breeding.,
Applied and Environmental Microbiology, Vol.75, No.17, 5529-5535, 2009.

(要約): Gene manipulation tools for an arachidonic-producing filamentous fungus, Mortierella alpina 1S-4, have not been sufficiently developed. In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was investigated for M. alpina 1S-4 transformation, using the uracil-auxotrophic mutant (ura5(-) strain) of M. alpina 1S-4 as a host strain and the homologous ura5 gene as a selectable marker gene. Furthermore, the gene for omega3-desaturase, catalyzing the conversion of n-6 fatty acid to n-3 fatty acid, was overexpressed in M. alpina 1S-4 by employing the ATMT system. As a result, we revealed that the frequency of transformation surpassed 400 transformants/10(8) spores, most of the integrated T-DNA appeared as a single copy at a random position in chromosomal DNA, and most of the transformants (60 to 80%) showed mitotic stability. Moreover, the accumulation of n-3 fatty acid in transformants was observed under the conditions of optimal omega3-desaturase gene expression. In particular, eicosapentaenoic acid (20:5n-3), an end product of n-3 fatty acids synthesized in M. alpina 1S-4, reached a maximum of 40% of total fatty acids. In conclusion, the ATMT system was found to be effective and suitable for the industrial strain Mortierella alpina 1S-4 and will be a useful tool for basic mutagenesis research and for industrial breeding of this strain.
(キーワード): Agrobacterium tumefaciens / Chromosomes, Fungal / Eicosapentaenoic Acid / Fatty Acid Desaturases / Fatty Acids / Gene Dosage / Gene Transfer Techniques / Genetic Engineering / Genomic Instability / Mortierella / Recombination, Genetic / Transformation, Genetic
(出版サイトへのリンク): ● Publication site (DOI): 10.1128/AEM.00648-09
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19581481; ● Search Scopus @ Elsevier (PMID): 19581481; ● Search Scopus @ Elsevier (DOI): 10.1128/AEM.00648-09

(DOI: 10.1128/AEM.00648-09, PubMed: 19581481) Masayoshi Muramatsu, Chikara Ohto, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Alkaline pH enhances farnesol production by Saccharomyces cerevisiae.,
Journal of Bioscience and Bioengineering, Vol.108, No.1, 52-55, 2009.

(要約): External environments affect prenyl alcohol production by squalene synthetase-deficient mutant Saccharomyces cerevisiae ATCC 64031. Cultivation of the yeast in medium with an initial pH ranging from 7.0 to 8.0 increased the amount of secreted farnesol (FOH). In contrast, acidic medium with a pH below 4.0 increased the intracellular FOH and its isomer nerolidol. These effects of alkaline pH were also observed on constant pH cultivation in a jar fermenter. On cultivation for 133 h, the FOH production reached 102.8 mg/l.
(キーワード): Culture Media / Farnesol / Fermentation / Hydrogen-Ion Concentration / Saccharomyces cerevisiae
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2009.02.012
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19577192; ● Search Scopus @ Elsevier (PMID): 19577192; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2009.02.012

(DOI: 10.1016/j.jbiosc.2009.02.012, PubMed: 19577192) Akinori Ando, Eiji Sakuradani, Kota Horinaka, Jun Ogawa and Sakayu Shimizu :
Transformation of an oleaginous zygomycete Mortierella alpina 1S-4 with the carboxin resistance gene conferred by mutation of the iron-sulfur subunit of succinate dehydrogenase.,
Current Genetics, Vol.55, No.3, 349-356, 2009.

(要約): The sdhB gene encoding an iron-sulfur (Ip) subunit of succinate dehydrogenase (SDH, EC 1.3.99.1) complex was cloned from Mortierella alpina 1S-4. The deduced amino acid sequence of SdhB from M. alpina 1S-4 showed high similarity to those of SdhB from other organisms. The mutated sdhB (CBXB) gene encodes a modified SdhB with an amino-acid substitution (a highly conserved histidine residue within the third cysteine-rich cluster of SdhB replaced by a leucine residue) and is known to confer carboxin resistance. We succeeded in transforming M. alpina 1S-4 by using the CBXB gene as a selectable marker gene and expressing the heterologous uidA gene encoding beta-glucuronidase of Escherichia coli. Moreover, transformation efficiency was up to 40-50 transformants per 4.0 x 10(8) spores. This carboxin-transformation system, characterized by marginal background growth and mitotic stability in M. alpina 1S-4, is considered to be widely useful for the wild strain, M. alpina 1S-4, and various derivative mutants without laborious preparation of auxotrophic mutants as a host strain.
(キーワード): Amino Acid Sequence / Blotting, Southern / Carboxin / DNA, Fungal / Drug Resistance, Fungal / Fungicides, Industrial / Glucuronidase / Molecular Sequence Data / Mortierella / Mutation / Protein Subunits / Sequence Analysis, DNA / Sequence Homology, Amino Acid / Succinate Dehydrogenase / Transformation, Genetic
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00294-009-0250-1
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19466416; ● Search Scopus @ Elsevier (PMID): 19466416; ● Search Scopus @ Elsevier (DOI): 10.1007/s00294-009-0250-1

(DOI: 10.1007/s00294-009-0250-1, PubMed: 19466416) Eiji Sakuradani, Masutoshi Nojiri, Haruna Suzuki and Sakayu Shimizu :
Identification of a novel fatty acid elongase with a wide substrate specificity from arachidonic acid-producing fungus Mortierella alpina 1S-4.,
Applied Microbiology and Biotechnology, Vol.84, No.4, 709-716, 2009.

(要約): The isolation and characterization of a gene (MALCE1) that encodes a fatty acid elongase from arachidonic acid-producing fungus Mortierella alpina 1S-4 are described. MALCE1 was confirmed to encode a fatty acid elongase by its expression in yeast Saccharomyces cerevisiae, resulting in the accumulation of 18-, 19-, and 20-carbon monounsaturated fatty acids and eicosanoic acid. Furthermore, the MALCE1 yeast transformant efficiently elongated exogenous 9-hexadecenoic acid, 9,12-octadecadienoic acid, and 9,12,15-octadecatrienoic acid. The MALCE1 gene-silenced strain obtained from M. alpina 1S-4 exhibited a low content of octadecanoic acid and a high content of hexadecanoic acid, compared with those in the wild strain. The enzyme encoded by MALCE1 was demonstrated to be involved in the conversion of hexadecanoic acid to octadecanoic acid, its main role in M. alpina 1S-4.
(キーワード): Acetyltransferases / Amino Acid Sequence / Arachidonic Acid / Cloning, Molecular / DNA, Fungal / Eicosanoic Acids / Gene Expression / Gene Silencing / Molecular Sequence Data / Mortierella / Palmitic Acid / Saccharomyces cerevisiae / Sequence Alignment / Sequence Analysis, DNA / Stearic Acids / Substrate Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-009-1999-3
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19415268; ● Search Scopus @ Elsevier (PMID): 19415268; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-009-1999-3

(DOI: 10.1007/s00253-009-1999-3, PubMed: 19415268) Akinori Ando, Jun Ogawa, Shigenobu Kishino, Taiyo Ito, Norifumi Shirasaka, Eiji Sakuradani, Kenzo Yokozeki and Sakayu Shimizu :
Fatty acid desaturation and elongation reactions of Trichoderma sp. 1-OH-2-3,
Journal of the American Oil Chemists' Society, Vol.86, No.3, 227-233, 2009.

(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s11746-008-1339-2
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1007/s11746-008-1339-2

(DOI: 10.1007/s11746-008-1339-2) Akinori Ando, Jun Ogawa, Satoshi Sugimoto, Shigenobu Kishino, Eiji Sakuradani, Kenzo Yokozeki and Sakayu Shimizu :
Selective production of cis-9,trans-11 isomer of conjugated linoleic acid from trans-vaccenic acid methyl ester by Delacroixia coronata.,
Journal of Applied Microbiology, Vol.106, No.5, 1697-1704, 2009.

(要約): Isomer selective bio-process for the practical production of cis-9,trans-11-CLA was first established. The process is benefitable for expanding the application of CLA for medicinal and nutraceutical purposes.
(キーワード): Culture Media / Fungi / Hydrogen-Ion Concentration / Isomerism / Linoleic Acids, Conjugated / Oleic Acids / Temperature / Time Factors
(出版サイトへのリンク): ● Publication site (DOI): 10.1111/j.1365-2672.2008.04139.x
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19226396; ● Search Scopus @ Elsevier (PMID): 19226396; ● Search Scopus @ Elsevier (DOI): 10.1111/j.1365-2672.2008.04139.x

(DOI: 10.1111/j.1365-2672.2008.04139.x, PubMed: 19226396) Eiji Sakuradani, Takahiro Abe, Kenji Matsumura, Akiko Tomi and Sakayu Shimizu :
Identification of mutation sites on Delta12 desaturase genes from Mortierella alpina 1S-4 mutants.,
Journal of Bioscience and Bioengineering, Vol.107, No.2, 99-101, 2009.

(要約): The mutation sites on the Delta12 desaturase gene in Mortierella alpina Delta12 desaturase-defective mutants SR88, TM912, and Mut48 accumulating Mead acid were identified. Each mutation resulted in an amino acid replacement (H116Y and P166L) in the Delta12 desaturase gene from SR88 and Mut48, respectively.
(キーワード): Base Sequence / DNA Primers / Fatty Acid Desaturases / Mortierella / Mutation
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2008.10.011
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19217543; ● Search Scopus @ Elsevier (PMID): 19217543; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2008.10.011

(DOI: 10.1016/j.jbiosc.2008.10.011, PubMed: 19217543) Chikara Ohto, Masayoshi Muramatsu, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Prenyl alcohol production by expression of exogenous isopentenyl diphosphate isomerase and farnesyl diphosphate synthase genes in Escherichia coli.,
Bioscience, Biotechnology, and Biochemistry, Vol.73, No.1, 186-188, 2009.

(要約): Isopentenyl diphosphate isomerase (idi) and farnesyl diphosphate synthase (ispA) genes were overexpressed in Escherichia coli. The resulting transformant showed 6.8-fold higher production of farnesol (389 microg/l). In a similar manner, overexpression of idi and mutated ispA led to high production of geranylgeraniol (128 microg/l).
(キーワード): Carbon-Carbon Double Bond Isomerases / Diterpenes / Escherichia coli / Escherichia coli Proteins / Farnesol / Geranyltranstransferase
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.80446
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19129660; ● Search Scopus @ Elsevier (PMID): 19129660; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.80446

(DOI: 10.1271/bbb.80446, PubMed: 19129660) Eiji Sakuradani, Takahiro Abe and Sakayu Shimizu :
Identification of mutation sites on omega3 desaturase genes from Mortierella alpina 1S-4 mutants.,
Journal of Bioscience and Bioengineering, Vol.107, No.1, 7-9, 2009.

(要約): The mutation sites on omega3 desaturase genes in two omega3 desaturase-defective mutants derived from arachidonic acid-producing Mortierella alpina 1S-4 were identified. The mutations each resulted in an amino acid replacement (W232Stop or W386Stop) which caused a lack of omega3 desaturase activity in these mutants.
(キーワード): Arachidonic Acid / DNA Mutational Analysis / Fatty Acid Desaturases / Fatty Acids / Fatty Acids, Unsaturated / Isoenzymes / Models, Biological / Models, Genetic / Mortierella / Mutation / Temperature / Time Factors
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2008.08.001
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19147101; ● Search Scopus @ Elsevier (PMID): 19147101; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2008.08.001

(DOI: 10.1016/j.jbiosc.2008.08.001, PubMed: 19147101) Chikara Ohto, Masayoshi Muramatsu, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Overexpression of the gene encoding HMG-CoA reductase in Saccharomyces cerevisiae for production of prenyl alcohols.,
Applied Microbiology and Biotechnology, Vol.82, No.5, 837-845, 2008.

(要約): To develop microbial production method for prenyl alcohols (e.g., (E,E)-farnesol (FOH), (E)-nerolidol (NOH), and (E,E,E)-geranylgeraniol (GGOH)), the genes encoding enzymes in the mevalonate and prenyl diphosphate pathways were overexpressed in Saccharomyces cerevisiae, and the resultant transformants were evaluated as to the production of these alcohols. Overexpression of the gene encoding hydroxymethylglutaryl (HMG)-CoA reductase was most effective among the genes tested. A derivative of S. cerevisiae ATCC 200589, which was selected through screening, was found to be the most suitable host for the production. On cultivation of the resultant transformant, in which the HMG-CoA reductase gene was overexpressed, in a 5-liter bench-scale jar fermenter for 7 d, the production of FOH, NOH, and GGOH reached 145.7, 98.8, and 2.46 mg/l, respectively.
(キーワード): Bioreactors / Fatty Alcohols / Fungal Proteins / Hydroxymethylglutaryl CoA Reductases / Industrial Microbiology / Metabolic Networks and Pathways / Mevalonic Acid / Promoter Regions, Genetic / Protein Structure, Tertiary / Recombinant Proteins / Saccharomyces cerevisiae / Terpenes
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-008-1807-5
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19083230; ● Search Scopus @ Elsevier (PMID): 19083230; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-008-1807-5

(DOI: 10.1007/s00253-008-1807-5, PubMed: 19083230) Eiji Sakuradani, Shoichi Murata, Hiroyuki Kanamaru and Sakayu Shimizu :
Functional analysis of a fatty acid elongase from arachidonic acid-producing Mortierella alpina 1S-4.,
Applied Microbiology and Biotechnology, Vol.81, No.3, 497-503, 2008.

(要約): We describe the isolation and characterization of a gene (MAELO) that encodes a fatty acid elongase from arachidonic acid-producing fungus Mortierella alpina 1S-4. Although the homologous MAELO gene had already been isolated from M. alpina ATCC 32221, its function had not yet been identified. The MAELO gene from M. alpina 1S-4 was confirmed to encode a fatty acid elongase by its expression in yeast Saccharomyces cerevisiae. Analysis of the fatty acid composition of the yeast transformant revealed the accumulation of 22-, 24-, and 26-carbon saturated fatty acids. On the other hand, RNA interference of the MAELO gene in M. alpina 1S-4 was carried out. The gene-silenced strain obtained on RNA interference exhibited low contents of 20-, 22-, and 24-carbon saturated fatty acids and a high content of stearic acid (18 carbons), compared with those in the wild strain. The enzyme encoded by the MAELO gene was demonstrated to be involved in the biosynthesis of 20-, 22-, and 24-carbon saturated fatty acids in M. alpina 1S-4.
(キーワード): Acetyltransferases / Amino Acid Sequence / Arachidonic Acid / Fatty Acids / Fungal Proteins / Gene Expression / Molecular Sequence Data / Mortierella / RNA Interference / Saccharomyces cerevisiae / Sequence Homology, Amino Acid
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-008-1675-z
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 18795285; ● Search Scopus @ Elsevier (PMID): 18795285; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-008-1675-z

(DOI: 10.1007/s00253-008-1675-z, PubMed: 18795285) Masayoshi Muramatsu, Chikara Ohto, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Various oils and detergents enhance the microbial production of farnesol and related prenyl alcohols.,
Journal of Bioscience and Bioengineering, Vol.106, No.3, 263-267, 2008.

(要約): The object of this research was improvement of prenyl alcohol production with squalene synthase-deficient mutant Saccharomyces cerevisiae ATCC 64031. On screening of many kinds of additives, we found that oils and detergents significantly enhanced the extracellular production of prenyl alcohols. Soybean oil showed the most prominent effect among the additives tested. Its effect was accelerated by a high concentration of glucose in the medium. The combination of these cultivation conditions led to the production of more than 28 mg/l of farnesol in the soluble fraction of the broth. The addition of these compounds to the medium was an effective method for large-scale production of prenyl alcohols with microorganisms.
(キーワード): Detergents / Farnesol / Farnesyl-Diphosphate Farnesyltransferase / Mutation / Neoprene / Plant Oils / Saccharomyces cerevisiae / Species Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1263/jbb.106.263
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 18930003; ● Search Scopus @ Elsevier (PMID): 18930003; ● Search Scopus @ Elsevier (DOI): 10.1263/jbb.106.263

(DOI: 10.1263/jbb.106.263, PubMed: 18930003) Masayoshi Muramatsu, Chikara Ohto, Shusei Obata, Eiji Sakuradani and Sakayu Shimizu :
Accumulation of prenyl alcohols by terpenoid biosynthesis inhibitors in various microorganisms.,
Applied Microbiology and Biotechnology, Vol.80, No.4, 589-595, 2008.

(要約): Squalene synthase inhibitors significantly accelerate the production of farnesol by various microorganisms. However, farnesol production by Saccharomyces cerevisiae ATCC 64031, in which the squalene synthase gene is deleted, was not affected by the inhibitors, indicating that farnesol accumulation is enhanced in the absence of squalene synthase activity. The combination of diphenylamine as an inhibitor of carotenoid biosynthesis and a squalene synthase inhibitor increases geranylgeraniol production by a yeast, Rhodotorula rubra NBRC 0870. An ent-kauren synthase inhibitor also enhances the production of farnesol and geranylgeraniol by a filamentous fungus, Gibberella fujikuroi NBRC 30336. These results indicate that the inhibition of downstream enzymes from prenyl diphosphate synthase leads to the production of farnesol and geranylgeraniol.
(キーワード): Alkyl and Aryl Transferases / Biosynthetic Pathways / Diphenylamine / Diterpenes / Enzyme Inhibitors / Farnesol / Farnesyl-Diphosphate Farnesyltransferase / Fungi / Terpenes
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-008-1578-z
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 18636253; ● Search Scopus @ Elsevier (PMID): 18636253; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-008-1578-z

(DOI: 10.1007/s00253-008-1578-z, PubMed: 18636253) Michihiko Kataoka, Takeru Ishige, Nobuyuki Urano, Yasushi Nakamura, Eiji Sakuradani, Satoko Fukui, Shinji Kita, Keiji Sakamoto and Sakayu Shimizu :
Cloning and expression of the L-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production.,
Applied Microbiology and Biotechnology, Vol.80, No.4, 597-604, 2008.

(要約): The gene encoding NADP(+)-dependent L-1-amino-2-propanol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 was cloned and sequenced. A 780-bp nucleotide fragment was confirmed to be the gene encoding AADH by agreement of the N-terminal and internal amino acid sequences of the purified AADH. The gene (aadh) codes a total of 259 amino acid residues, and the deduced amino acid sequence shows similarity to several short-chain dehydrogenase/reductase family proteins. An expression vector, pKKAADH, which contains the full length aadh was constructed. Escherichia coli cells possessing pKKAADH exhibited a 10.4-fold increase in specific activity as to catalysis of the reduction of (S)-1-phenyl-2-methylaminopropan-1-one (MAK), as compared with that of R. erythropolis MAK154 induced by 1-amino-2-propanol (1 mg/ml). Coexpression of aadh with a cofactor regeneration enzyme (glucose dehydrogenase) gene was also performed, and a system for sufficient production of d-pseudoephedrine from racemic MAK was constructed.
(キーワード): Adamantane / Alcohol Oxidoreductases / Bacterial Proteins / Cloning, Molecular / Escherichia coli / Gene Expression / Glucose 1-Dehydrogenase / Molecular Sequence Data / Rhodococcus
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-008-1563-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 18584170; ● Search Scopus @ Elsevier (PMID): 18584170; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-008-1563-6

(DOI: 10.1007/s00253-008-1563-6, PubMed: 18584170) Hiroki Ukitsu, Takashi Kuromori, Kiminori Toyooka, Yumi Goto, Ken Matsuoka, Eiji Sakuradani, Sakayu Shimizu, Asako Kamiya, Yuko Imura, Masahiro Yuguchi, Takuji Wada, Takashi Hirayama and Kazuo Shinozaki :
Cytological and biochemical analysis of COF1, an Arabidopsis mutant of an ABC transporter gene.,
Plant & Cell Physiology, Vol.48, No.11, 1524-1533, 2007.

(要約): In transposon-tagged lines of Arabidopsis we found two new mutants, cof1-1 and cof1-2 (cuticular defect and organ fusion), that show the phenotype of wilting when grown in soil, organ fusion of rosette leaves and infertility. Toluidine blue testing and scanning electron microscopy observation revealed that these mutants had cuticular defects in the stems and adult leaves, but not in cotyledones. Transmission electron microscopy observation revealed thinner cuticle layers in the mutants, and cuticular materials interspersed between the two fused epidermal layers were observed in the mutant rosette leaves. These two mutants had a transposon insertion in the coding regions of WBC11, which was classified as a member of ABC transporter genes in Arabidopsis. WBC11 showed high sequence similarity to CER5 (also called WBC12), which was involved in cuticular lipid export. Gas chromatographic analysis revealed that C29 alkane extracted from the stem surface of cof1 mutants was reduced whereas C29 ketone was accumulated, which was different from the case of cer5 mutants. While cer5 mutants had fairly normal morphology, cof1 mutants had pleiotropic phenotypes so that COF1/WBC11 could have important roles different from those of CER5/WBC12. We also found that C29 alkane was accumulated in the intracellular extract of cof1 mutants, suggesting a function for WBC11 in the direct transport of lipid molecules. Pollen observation showed that mutant pollen grains were irregularly shaped. The function of COF1/WBC11 in lipid transport for the construction of cuticle layers and pollen coats for normal organ formation is discussed.
(キーワード): ATP-Binding Cassette Transporters / Arabidopsis / Arabidopsis Proteins / Chromatography, Gas / Gene Expression Regulation, Plant / Luminescent Proteins / Microscopy, Confocal / Microscopy, Electron, Scanning / Microscopy, Electron, Transmission / Mutation / Phylogeny / Plant Epidermis / Plant Leaves / Plants, Genetically Modified / Pollen / Recombinant Fusion Proteins / Reverse Transcriptase Polymerase Chain Reaction
(出版サイトへのリンク): ● Publication site (DOI): 10.1093/pcp/pcm139
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 17971336; ● Search Scopus @ Elsevier (PMID): 17971336; ● Search Scopus @ Elsevier (DOI): 10.1093/pcp/pcm139

(DOI: 10.1093/pcp/pcm139, PubMed: 17971336) Shuo Zhang, Eiji Sakuradani and Sakayu Shimizu :
Identification of a sterol Delta7 reductase gene involved in desmosterol biosynthesis in Mortierella alpina 1S-4.,
Applied and Environmental Microbiology, Vol.73, No.6, 1736-1741, 2007.

(要約): Molecular cloning of the gene encoding sterol Delta7 reductase from the filamentous fungus Mortierella alpina 1S-4, which accumulates cholesta-5,24-dienol (desmosterol) as the main sterol, revealed that the open reading frame of this gene, designated MoDelta7SR, consists of 1,404 bp and codes for 468 amino acids with a molecular weight of 53,965. The predicted amino acid sequence of MoDelta7SR showed highest homology of 51% with that of sterol Delta7 reductase (EC 1.3.1.21) from Xenopus laevis (African clawed frog). Heterologous expression of the MoDelta7SR gene in yeast Saccharomyces cerevisiae revealed that MoDelta7SR converts ergosta-5,7-dienol to ergosta-5-enol (campesterol) by the activity of Delta7 reductase. In addition, with gene silencing of MoDelta7SR gene by RNA interference, the transformant accumulated cholesta-5,7,24-trienol up to 10% of the total sterols with a decrease in desmosterol. Cholesta-5,7,24-trienol is not detected in the control strain. This indicates that MoDelta7SR is involved in desmosterol biosynthesis in M. alpina 1S-4. This study is the first report on characterization of sterol Delta7 reductase from a microorganism.
(キーワード): Animals / Base Sequence / Cholecalciferol / Cholesterol / Cloning, Molecular / DNA, Fungal / Desmosterol / Ergosterol / Gas Chromatography-Mass Spectrometry / Gene Expression / Gene Silencing / Molecular Sequence Data / Molecular Structure / Molecular Weight / Mortierella / Open Reading Frames / Oxidoreductases Acting on CH-CH Group Donors / Phytosterols / Saccharomyces cerevisiae / Sequence Homology, Amino Acid / Sterols / Xenopus laevis
(出版サイトへのリンク): ● Publication site (DOI): 10.1128/AEM.02425-06
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 17220261; ● Search Scopus @ Elsevier (PMID): 17220261; ● Search Scopus @ Elsevier (DOI): 10.1128/AEM.02425-06

(DOI: 10.1128/AEM.02425-06, PubMed: 17220261) Shuo Zhang, Eiji Sakuradani, Kuni Ito and Sakayu Shimizu :
Identification of a novel bifunctional delta12/delta15 fatty acid desaturase from a basidiomycete, Coprinus cinereus TD#822-2.,
FEBS Letters, Vol.581, No.2, 315-319, 2007.

(要約): A new gene encoding a delta12 fatty acid desaturase-related protein was cloned from a multicellular basidiomycete Coprinus cinereus TD#822-2. The 1326 bp full-length gene, designated as Cop-odeA, codes for a putative protein of 442 amino acids with a MW of 49224. The Cop-odeA yeast transformant accumulated four new fatty acids identified as 9,12-hexadecadienoic acid, 9,12,15-hexadecatrienoic acid, linoleic acid, and alpha-linolenic acid, which comprised 8.8%, 1.0%, 29.0%, and 0.6% of the total fatty acids, respectively. The Cop-odeA protein was confirmed to be a novel bifunctional fatty acid desaturase with both high delta12 desaturase activity and unusual delta15 desaturase activity.
(キーワード): Cloning, Molecular / Coprinus / Fatty Acid Desaturases / Fatty Acids / Phylogeny / Saccharomyces cerevisiae
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.febslet.2006.12.031
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 17208234; ● Search Scopus @ Elsevier (PMID): 17208234; ● Search Scopus @ Elsevier (DOI): 10.1016/j.febslet.2006.12.031

(DOI: 10.1016/j.febslet.2006.12.031, PubMed: 17208234) Yuzo Kojima, Eiji Sakuradani and Sakayu Shimizu :
Acidolysis and glyceride synthesis reactions using fatty acids with two Pseudomonas lipases having different substrate specificities.,
Journal of Bioscience and Bioengineering, Vol.102, No.3, 179-183, 2006.

(要約): Enzymatic acidolysis and glyceride synthesis using polyunsaturated fatty acids (PUFAs) with lipases from Pseudomonas fluorescens HU380 (HU-lipase), P. fluorescens AK102 (AK-lipase), and Candida rugosa (CR-lipase) were studied. The acidolysis of triolein with eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) in n-hexane was evaluated with lipases immobilized on Celite 545. HU-lipase showed the highest incorporation rate at a low temperature (10 degrees C) with either EPA or DHA as the acyl donor, and the rate decreased with increasing reaction temperature. At 45 degrees C, the rates for EPA and DHA were 7.1 and 0.5 relative to those at 10 degrees C, respectively. The EPA incorporation rate was even higher at a low temperature (10 degrees C), and the DHA incorporation rate increased with decreasing temperature. Although AK-lipase showed the reverse tendency for incorporation rate, the DHA incorporation rate increased with increasing reaction temperature with both PUFAs. HU-lipase reacted well with PUFAs such as DHA, EPA, arachidonic acid (AA), mead acid (MA), and dihomo-gamma-linolenic acid (DGLA) on acidolysis and glyceride synthesis. The reactivities of AK-lipase toward these PUFAs except for DGLA, i.e., MA, AA, EPA, and DHA, were low for both reactions. The unique substrate specificities of the lipases from the Pseudomonas strains will enable us to use these lipases for the modification of fats and oils containing PUFAs such as fish oil.
(キーワード): Bacterial Proteins / Candida / Cold Temperature / Fatty Acids / Fungal Proteins / Glycerides / Lipase / Pseudomonas fluorescens / Substrate Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1263/jbb.102.179
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 17046530; ● Search Scopus @ Elsevier (PMID): 17046530; ● Search Scopus @ Elsevier (DOI): 10.1263/jbb.102.179

(DOI: 10.1263/jbb.102.179, PubMed: 17046530) Yuzo Kojima, Eiji Sakuradani and Sakayu Shimizu :
Different specificity of two types of Pseudomonas lipases for C20 fatty acids with a Delta5 unsaturated double bond and their application for selective concentration of fatty acids.,
Journal of Bioscience and Bioengineering, Vol.101, No.6, 496-500, 2006.

(要約): Two kinds of lipases, AK-lipase and HU-lipase, produced by two different Pseudomonas fluorescens strains, AK102 and HU380, respectively, were evaluated as to fatty acid hydrolysis specificity using six types of oil containing higher amounts of C20 fatty acids such as arachidonic acid (5,8,11,14-eicosatetraenoic acid, AA, or 20:4omega6), dihomo-gamma-linolenic acid (8,11,14-eicosatrienoic acid, DGLA, or 20:3omega6), 5,8,11,14,17-eicosapentaenoic acid (EPA or 20:5omega3), mead acid (5,8,11-eicosatrienoic acid, MA, or 20:3omega9), 8,11-eicosadienoic acid (20:2omega9) and 8,11,14,17-eicosatetraenoic acid (20:4omega3). Although HU-lipase did not show any specificity for C20 fatty acids with respect to the presence or absence of a Delta5 unsaturated bond, it exhibited comparatively low reactivity for 4,7,10,13,16,19-docosahexaenoic acid (DHA or 22:6omega3). In contrast, AK-lipase was less reactive for C20 fatty acids with a Delta5 unsaturated bond. However, the specificity of hydrolysis of AK-lipase gradually decreased as the reaction proceeded. Utilizing this fatty acid specificity, we concentrated either EPA or DHA from fish oils containing both EPA and DHA by means of lipase-catalyzed hydrolysis and urea adduction. Hydrolysis and urea adduction of refined cod oil including 12.2% EPA and 6.9% DHA with HU-lipase provided free fatty acids with 43.1% EPA and 7% DHA, respectively. The resulting yield of concentrated total fatty acids comprised 2.6% of the fatty acids from the cod oil. Thus, EPA was particularly concentrated in the fatty acids derived from refined cod oil on partial hydrolysis with HU-lipase followed by urea adduction. On the other hand, hydrolysis of cuttlefish oil with AK-lipase followed by urea adduction increase slightly the EPA composition from 14.2% to 16.8%, and markedly enhanced the composition of DHA from 16.3% to 44.6% in the hydrolyzed fatty acids. The yield of purified total fatty acids by urea concentrate was 9.4% of the fatty acids from the cuttlefish oil. Thus, DHA was particularly concentrated in the fatty acids derived from on partial hydrolysis with AK-lipase followed by urea adduction. We concluded that EPA and DHA concentrates can be easily and inexpensively obtained using HU-lipase and AK-lipase, respectively. Furthermore, it might be possible to separate and concentrate C20 polyunsaturated fatty acids (PUFAs) with or without a Delta5 double bond from PUFAs rich oils including both fatty acids.
(キーワード): Chemical Fractionation / Docosahexaenoic Acids / Eicosapentaenoic Acid / Enzyme Activation / Fatty Acids, Unsaturated / Lipase / Pseudomonas fluorescens / Substrate Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1263/jbb.101.496
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16935251; ● Search Scopus @ Elsevier (PMID): 16935251; ● Search Scopus @ Elsevier (DOI): 10.1263/jbb.101.496

(DOI: 10.1263/jbb.101.496, PubMed: 16935251) Shuo Zhang, Eiji Sakuradani and Sakayu Shimizu :
Identification and production of n-8 odd-numbered polyunsaturated fatty acids by a delta12 desaturation-defective mutant of Mortierella alpina 1S-4.,
Lipids, Vol.41, No.6, 623-626, 2006.

(要約): A mutant of Mortierella alpina, JT-180, which is defective in its delta12 desaturase activity but exhibits enhanced activities of delta5 and delta6 desaturases, produced a high proportion (up to 80%) of odd-chain FA when grown on 3% n-heptadecane. Following growth of the mutant on n-heptadecane, three unusual odd-chain fatty acyl residues were identified as 6,9-heptadecadienoic acid (17:2), 8,11 -nonadecadienoic acid (19:2), and 5,8,11-nonadecatrienoic acid (19:3) by means of GC-MS, MS-MS, and NMR analyses. The mycelial contents of these FA reached 20.3, 3.6, and 5.8 mg/g dry mycelia, respectively, when it was cultivated in medium comprising 4% (vol/vol) n-heptadecane and 1% (wt/vol) yeast extract, pH 6.0, at 28 degrees C for 7 d. The biosynthetic route (n-8 route) to 19:3 was presumed to mimic the n-9 route to Mead acid (20:3n-9) in mammals: 17:0 --> 9-17:1 --> 6,9-17:2 --> 8,11-19:2 --> 5,8,11-19:3.
(キーワード): Alkanes / Cell Culture Techniques / Fatty Acid Desaturases / Fatty Acids, Unsaturated / Fungi / Lipids / Models, Biological / Mortierella
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s11745-006-5011-z
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16981440; ● Search Scopus @ Elsevier (PMID): 16981440; ● Search Scopus @ Elsevier (DOI): 10.1007/s11745-006-5011-z

(DOI: 10.1007/s11745-006-5011-z, PubMed: 16981440) Ren Chen, Keisuke Matsui, Miyuki Ogawa, Mieko Oe, Misa Ochiai, Hiroshi Kawashima, Eiji Sakuradani, Sakayu Shimizu, Masao Ishimoto, Makoto Hayashi, Yoshikatsu Murooka and Yoshikazu Tanaka :
Expression of Δ6, Δ5 desaturase and GLELO elongase genes from Mortierella alpina for production of arachidonic acid in soybean [Glycine max (L.) Merrill] seeds,
Plant Science, Vol.170, No.2, 399-406, 2006.

(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.plantsci.2005.09.006
(文献検索サイトへのリンク): ● Summary page in Scopus @ Elsevier: 2-s2.0-29244437851

(DOI: 10.1016/j.plantsci.2005.09.006, Elsevier: Scopus) Ken-ichi Hashimoto, Hisashi Kawasaki, Kouki Akazawa, Jun Nakamura, Youko Asakura, Takuji Kudo, Eiji Sakuradani, Sakayu Shimizu and Tsuyoshi Nakamatsu :
Changes in composition and content of mycolic acids in glutamate-overproducing Corynebacterium glutamicum.,
Bioscience, Biotechnology, and Biochemistry, Vol.70, No.1, 22-30, 2006.

(要約): Overproduction of glutamate by Corynebacterium glutamicum is induced by biotin limitation or by the supplementation of specific detergents, sublethal amounts of penicillin, or cerulenin. But, it remains unclear why these different treatments, which have different sites of primary action, produce similar effects. In this study, it was found that the cellular content of mycolic acids--characteristic constituents of Corynebacterineae that are synthesized from fatty acids and form a cell surface layer--decreased under all conditions that induced glutamate overproduction. Furthermore, short mycolic acids increased under conditions of biotin limitation and cerulenin supplementation. These results suggest that different treatments produce the same effect that causes defects in the mycolic acid layer. This is perhaps one of the key factors in overproduction of glutamate by C. glutamicum.
(キーワード): Cerulenin / Chromatography, Gas / Corynebacterium glutamicum / Esters / Glutamates / Mass Spectrometry / Methylation / Molecular Structure / Mycolic Acids
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.70.22
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16428817; ● Search Scopus @ Elsevier (PMID): 16428817; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.70.22

(DOI: 10.1271/bbb.70.22, PubMed: 16428817) Seiki Takeno, Eiji Sakuradani, Akiko Tomi, Misa Inohara-Ochiai, Hiroshi Kawashima and Sakayu Shimizu :
Transformation of oil-producing fungus, Mortierella alpina 1S-4, using Zeocin, and application to arachidonic acid production.,
Journal of Bioscience and Bioengineering, Vol.100, No.6, 617-622, 2005.

(要約): The arachidonic acid-producing fungus Mortierella alpina 1S-4, an industrial strain, was endowed with Zeocin resistance by integration of the Zeocin-resistance gene at the rDNA locus of genomic DNA. Plasmid DNA was introduced into spores by microprojectile bombardment. Twenty mg/ml Zeocin completely inhibited the germination of M. alpina 1S-4 spores, and decreased the growth rate of fungal filaments to some extent. It was suggested that preincubation period and temperature had a great influence on transformation efficiency. Four out of 26 isolated transformants were selected. Molecular analysis of these stable transformants showed that the plasmid DNA was integrated into the rDNA locus of the genomic DNA. We expect that this system will be applied for useful oil production by gene manipulation of M. alpina 1S-4 and its derivative mutants. On the basis of the fundamental transformation system, we also tried to overexpress a homologous polyunsaturated fatty acid elongase gene, which has been reported to be included in the rate-limiting step for arachidonic acid production, thereby leading to increased arachidonic acid production.
(キーワード): Arachidonic Acid / Bleomycin / Cell Culture Techniques / Drug Resistance, Fungal / Genetic Enhancement / Mortierella / Transfection
(出版サイトへのリンク): ● Publication site (DOI): 10.1263/jbb.100.617
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16473770; ● Search Scopus @ Elsevier (PMID): 16473770; ● Search Scopus @ Elsevier (DOI): 10.1263/jbb.100.617

(DOI: 10.1263/jbb.100.617, PubMed: 16473770) Ren Chen, Shinzo Tsuda, Keisuke Matsui, Masako Fukuchi-Mizutani, Misa Ochiai, Sakayu Shimizu, Eiji Sakuradani, Toshio Aoki, Ryujiro Imaizumi, Shin-ichi Ayabe and Yoshikazu Tanaka :
Production of γ-linolenic acid in Lotus japonicus and Vigna angularis by expression of the Δ6-fatty-acid desaturase gene isolated from Mortierella alpina,
Plant Science, Vol.169, No.3, 599-605, 2005.

(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.plantsci.2005.05.005
(文献検索サイトへのリンク): ● Summary page in Scopus @ Elsevier: 2-s2.0-22444439943

(DOI: 10.1016/j.plantsci.2005.05.005, Elsevier: Scopus) Seiki Takeno, Eiji Sakuradani, Akiko Tomi, Misa Inohara-Ochiai, Hiroshi Kawashima, Toshihiko Ashikari and Sakayu Shimizu :
Improvement of the fatty acid composition of an oil-producing filamentous fungus, Mortierella alpina 1S-4, through RNA interference with delta12-desaturase gene expression.,
Applied and Environmental Microbiology, Vol.71, No.9, 5124-5128, 2005.

(要約): An oleaginous fungus, Mortierella alpina 1S-4, is used commercially for arachidonic acid production. Delta12-Desaturase, which desaturates oleic acid (18:1n-9) to linoleic acid (18:2n-6), is a key enzyme in the arachidonic acid biosynthetic pathway. To determine if RNA interference (RNAi) by double-stranded RNA occurs in M. alpina 1S-4, we silenced the Delta12-desaturase gene. The silenced strains accumulate 18:2n-9, 20:2n-9, and Mead acid (20:3n-9), which are not detected in either the control strain or wild type strain 1S-4. The fatty acid composition of stable transformants was similar to that of Delta12-desaturation-defective mutants previously identified. Thus, RNAi occurs in M. alpina and could be used to alter the types and relative amounts of fatty acids produced by commercial strains of this fungus without mutagenesis or other permanent changes in the genetic background of the producing strains.
(キーワード): Arachidonic Acid / Culture Media / Fatty Acid Desaturases / Fatty Acids, Unsaturated / Gene Silencing / Industrial Microbiology / Mortierella / RNA Interference / RNA, Fungal / Transformation, Genetic
(出版サイトへのリンク): ● Publication site (DOI): 10.1128/AEM.71.9.5124-5128.2005
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16151095; ● Search Scopus @ Elsevier (PMID): 16151095; ● Search Scopus @ Elsevier (DOI): 10.1128/AEM.71.9.5124-5128.2005

(DOI: 10.1128/AEM.71.9.5124-5128.2005, PubMed: 16151095) Takahiro Abe, Eiji Sakuradani, Takahiro Asano, Hiroyuki Kanamaru and Sakayu Shimizu :
Functional characterization of Delta9 and omega9 desaturase genes in Mortierella alpina 1S-4 and its derivative mutants.,
Applied Microbiology and Biotechnology, Vol.70, No.6, 711-719, 2005.

(要約): Cloning and characterization of the Delta9 desaturase (Delta9I) gene of a fungus, Mortierella alpina 1S-4, was previously reported. In this study, two genes encoding Delta9 desaturase homologs were isolated from this fungus. One is a Delta9 desaturase (Delta9II) that exhibits 86% amino acid sequence similarity to Delta9I. Functional analysis involving expression of the encoding gene in Aspergillus oryzae revealed that Delta9II exhibits Delta9 desaturase activity, 18:0 being converted to 18:1Delta9. However, unlike Delta9I, the Delta9II transformant accumulated a low amount of 16:1Delta9. The other homolog is a omega9 desaturase (omega9) that exhibits 56 and 58% amino acid sequence similarity to Delta9I and Delta9II, respectively. On functional analysis with the Aspergillus transformant, it was found that omega9 does not convert 18:0 to 18:1Delta9, but converts 24:0 and 26:0 to 24:1omega9 and 26:1omega9, respectively. On the other hand, Delta9 desaturation-defective mutants characterized by accumulation of 18:0 were derived from M. alpina 1S-4 with a chemical mutagen, and the mutated sites of the Delta9 desaturase genes were identified. The mutation on the Delta9I gene was assumed to cause an amino acid replacement (W136Stop, G265D, and W360Stop) in the mutants (HR222, T4, and ST56), respectively. In these mutants, there was no mutated site on the Delta9II and omega9 genes. Real-time quantitative PCR (RTQ-PCR) analysis revealed that (1) the transcriptional level of the Delta9I gene in HR222 and T4 was much higher than that in the wild strain until the fifth day of the cultivation periods, (2) the Delta9II gene of the mutants was transcribed until the fourth day at the same level as the Delta9I gene of the wild strain, whereas the Delta9II gene of the wild strain was transcribed at a lower level, and (3) the transcriptional level of the omega9 gene in both the mutants and the wild strain was low, i.e., as low as that of the Delta9II gene of the wild strain. In these Delta9 desaturation-defective mutants, Delta9II is likely to play an important role in Delta9 desaturation.
(キーワード): Amino Acid Sequence / Aspergillus oryzae / Cloning, Molecular / Fatty Acid Desaturases / Fatty Acids / Molecular Sequence Data / Mortierella / Mutation / Sequence Analysis, DNA
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-005-0115-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16133334; ● Search Scopus @ Elsevier (PMID): 16133334; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-005-0115-6

(DOI: 10.1007/s00253-005-0115-6, PubMed: 16133334) Takahiro Abe, Eiji Sakuradani, Takahiro Asano, Hiroyuki Kanamaru, Yuji Ioka and Sakayu Shimizu :
Identification of mutation sites on delta6 desaturase genes from Mortierella alpina 1S-4 mutants.,
Bioscience, Biotechnology, and Biochemistry, Vol.69, No.5, 1021-1024, 2005.

(要約): Three Delta6 desaturase-defective mutants, designated YB214, HR95, and ST66, were newly isolated from Mortierella alpina 1S-4, after treating wild-type spores with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). These three mutants and Mut49, isolated previously, are capable of accumulating 5,11,14-cis-eicosatrienoic acid (20:3Delta5). Two functional Delta6 desaturases (Delta6I and Delta6II) were found to exist in M. alpina 1S-4. The mutation sites on the Delta6I gene in the Delta6 desaturase-defective mutants were identified. The mutations each resulted in an amino acid replacement (W314Stop, T375K, and G390D) in Delta6I from ST66, HR95, and YB214 respectively, and uncorrected transcription of the Delta6I gene in Mut49 was caused by disappearance of the GT-terminal of the second intron, resulting in low Delta6 desaturation activity in these mutants. On the other hand, there was no mutation site on the Delta6II genes of the mutants.
(キーワード): Amino Acid Sequence / Arachidonic Acids / Base Sequence / Fatty Acid Desaturases / Molecular Sequence Data / Mortierella / Mutation
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.69.1021
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15914925; ● Search Scopus @ Elsevier (PMID): 15914925; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.69.1021

(DOI: 10.1271/bbb.69.1021, PubMed: 15914925) Takahiro Abe, Eiji Sakuradani, Takuji Ueda and Sakayu Shimizu :
Identification of mutation sites on delta5 desaturase genes from Mortierella alpina 1S-4 mutants.,
Journal of Bioscience and Bioengineering, Vol.99, No.3, 296-299, 2005.

(要約): The mutation sites on delta5 desaturase genes in delta5 desaturase-defective mutants derived from arachidonic acid-producing Mortierella alpina 1S-4 were identified. The mutations resulted in an amino acid replacement (G189E or W301Stop) and uncorrected transcription caused by recognition of an AG-terminal newly created on C207A gene mutation, resulting in low or no delta5 desaturase activity in these mutants.
(キーワード): Amino Acid Sequence / Amino Acid Substitution / Base Sequence / DNA Mutational Analysis / Fatty Acid Desaturases / Molecular Sequence Data / Mortierella / Sequence Homology, Amino Acid / Structure-Activity Relationship
(出版サイトへのリンク): ● Publication site (DOI): 10.1263/jbb.99.296
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16233793; ● Search Scopus @ Elsevier (PMID): 16233793; ● Search Scopus @ Elsevier (DOI): 10.1263/jbb.99.296

(DOI: 10.1263/jbb.99.296, PubMed: 16233793) Seiki Takeno, Eiji Sakuradani, Shoichi Murata, Misa Inohara-Ochiai, Hiroshi Kawashima, Toshihiko Ashikari and Sakayu Shimizu :
Molecular evidence that the rate-limiting step for the biosynthesis of arachidonic acid in Mortierella alpina is at the level of an elongase.,
Lipids, Vol.40, No.1, 25-30, 2005.

(要約): The oil-producing fungus Mortierella alpina 1S-4 is an industrial strain for arachidonic acid (AA) production. To determine its physiological properties and to clarify the biosynthetic pathways for PUFA, heterologous and homologous gene expression systems were established in this fungus. The first trial was performed with an enhanced green fluorescent protein gene to assess the transformation efficiency for heterologous gene expression. As a result, strong fluorescence was observed in the spores of the obtained transformant, suggesting that the foreign gene was inherited by the spores. The next trial was performed with a homologous PUFA elongase (GLELOp) gene, this enzyme having been reported to catalyze the elongation of GLA (18:3n-6) to dihomo-gamma-linolenic acid (20:3n-6), and to be the rate-limiting step of AA production. The FA composition of the transformant was different from that of the host strain: The GLA content was decreased whereas that of AA was increased. These data support the hypothesis that the GLELOp enzyme plays an important role in PUFA synthesis, and may indicate how to control PUFA biosynthesis.
(キーワード): Acetyltransferases / Arachidonic Acid / Fatty Acids / Fatty Acids, Unsaturated / Fungal Proteins / Kinetics / Mortierella / gamma-Linolenic Acid
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s11745-005-1356-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15825827; ● Search Scopus @ Elsevier (PMID): 15825827; ● Search Scopus @ Elsevier (DOI): 10.1007/s11745-005-1356-6

(DOI: 10.1007/s11745-005-1356-6, PubMed: 15825827) Eiji Sakuradani, Yuriko Hirano, Nozomu Kamada, Masutoshi Nojiri, Jun Ogawa and Sakayu Shimizu :
Improvement of arachidonic acid production by mutants with lower n-3 desaturation activity derived from Mortierella alpina.,
Applied Microbiology and Biotechnology, Vol.66, No.3, 243-248, 2004.

(要約): Five mutants were obtained, Y11, Y135, Y164, Y180 and Y61, capable of accumulating higher amounts of arachidonic acid (AA) than Mortierella alpina 1S-4, an industrial strain for the production of AA-rich triacylglycerol (TG). This is thought to be due to low or no activity of n-3 desaturation with conversion of AA to eicosapentaenoic acid, which functions at a cultural temperature below 20 degrees C. In small-scale cultivation under optimum conditions, Y11 and Y61 respectively accumulated 4.97 mg/ml and 4.11 mg/ml of AA, using a high concentration of glucose at 20 degrees C, compared with 3.74 mg/ml for M. alpina 1S-4. In a 5-1 jar fermentor, the AA content in Y11 and Y61 kept increasing during cultivation, with consumption of the glucose in the medium; and this reached 1.48 mg/ml and 1.77 mg/ml (118 mg/g, 120 mg/g of dry mycelia) at day 10, respectively, compared with 0.95 mg/ml (86 mg/g of dry mycelia) for M. alpina 1S-4. From the results of lipid analysis, the TG contents of Y11 and Y61 in the major lipids were significantly higher than that of M. alpina 1S-4; and the AA percentages in TG of Y11 and Y61 were also higher. Both Y11 and Y61 are potential producers of TG rich in AA.
(キーワード): Arachidonic Acids / Fatty Acid Desaturases / Glucose / Mortierella / Mutation / Temperature / Triglycerides
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-004-1682-7
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15300418; ● Search Scopus @ Elsevier (PMID): 15300418; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-004-1682-7

(DOI: 10.1007/s00253-004-1682-7, PubMed: 15300418) Eiji Sakuradani, Takahiro Abe, Keita Iguchi and Sakayu Shimizu :
A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4.,
Applied Microbiology and Biotechnology, Vol.66, No.6, 648-654, 2004.

(要約): A filamentous fungus, Mortierella alpina 1S-4, is capable of producing not only arachidonic acid (AA; 20:4n-6) but also eicosapentaenoic acid (EPA; 20:5n-3) below a cultural temperature of 20 degrees C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel omega3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 Delta12-desaturase and Saccharomyces kluyveri omega3-desaturase, the omega3-desaturase gene from M. alpina 1S-4 was cloned. Homology analysis of protein databases revealed that the amino acid sequence showed 51% identity, at the highest, with M. alpina 1S-4 Delta12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri omega3-desaturase. The cloned cDNA was confirmed to encode the omega3-desaturase by its expression in the yeast Sac. cerevisiae. Analysis of the fatty acid composition of the yeast transformant demonstrated that 18-carbon and 20-carbon n-3 polyunsaturated fatty acids (PUFAs) were accumulated through conversion of exogenous 18-carbon and 20-carbon n-6 PUFAs. The substrate specificity of the M. alpina 1S-4 omega3-desaturase differs from those of the known fungal omega3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri omega3-desaturases desaturate 18-carbon n-6 PUFAs, Spr. diclina omega3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans omega3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 omega3-desaturase is rather similar to that of C. elegans omega3-desaturase, but the M. alpina omega3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 omega3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates.
(キーワード): Amino Acid Sequence / Animals / Caenorhabditis elegans / Cloning, Molecular / DNA, Fungal / Fatty Acid Desaturases / Fatty Acids / Fatty Acids, Unsaturated / Fungal Proteins / Genes, Fungal / Molecular Sequence Data / Mortierella / Phylogeny / Saccharomyces / Saprolegnia / Sequence Alignment / Sequence Analysis, DNA / Sequence Homology, Amino Acid / Substrate Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-004-1760-x
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15538555; ● Search Scopus @ Elsevier (PMID): 15538555; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-004-1760-x

(DOI: 10.1007/s00253-004-1760-x, PubMed: 15538555) Seiki Takeno, Eiji Sakuradani, Shoichi Murata, Misa Inohara-Ochiai, Hiroshi Kawashima, Toshihiko Ashikari and Sakayu Shimizu :
Establishment of an overall transformation system for an oil-producing filamentous fungus, Mortierella alpina 1S-4.,
Applied Microbiology and Biotechnology, Vol.65, No.4, 419-425, 2004.

(要約): Oil-producing fungus Mortierella alpina 1S-4 is an industrial strain. To determine its physiological properties and to clarify the biosynthetic pathways for polyunsaturated fatty acids, a transformation system for this fungus was established using a derivative of it, i.e., a ura5- mutant lacking orotate phosphoribosyl transferase (OPRTase, EC.2.4.2.10) activity. Transformation with a vector containing the homologous ura5 gene as a marker was successfully performed using microprojectile bombardment, other methods frequently used for transformation, such as the protoplasting, lithium acetate, or electroporation methods, not giving satisfactory results. As a result, two types of transformants were obtained: a few stable transformants overexpressing the ura5 gene, and many unstable transformants showing OPRTase activity comparable to that of the wild-type strain. The results of quantitative PCR indicated that the stable transformants could retain the ura5 genes originating from the transformation vector regardless of the culture conditions. On the other hand, unstable transformants easily lost the marker gene under uracil-containing conditions, as expected. In this paper, we report that an overall transformation system for this fungus was successfully established, and propose how to select useful transformants as experimental and industrial strains.
(キーワード): Acetates / Culture Media / Electroporation / Genes, Fungal / Genetic Vectors / Mortierella / Mutation / Oils / Orotate Phosphoribosyltransferase / Protoplasts / Selection, Genetic / Transformation, Genetic
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-004-1622-6
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15138730; ● Search Scopus @ Elsevier (PMID): 15138730; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-004-1622-6

(DOI: 10.1007/s00253-004-1622-6, PubMed: 15138730) Seiki Takeno, Eiji Sakuradani, Shoichi Murata, Misa Inohara-Ochiai, Hiroshi Kawashima, Toshihiko Ashikari and Sakayu Shimizu :
Cloning and sequencing of the ura3 and ura5 genes, and isolation and characterization of uracil auxotrophs of the fungus Mortierella alpina 1S-4.,
Bioscience, Biotechnology, and Biochemistry, Vol.68, No.2, 277-285, 2004.

(要約): The oil-producing fungus Mortierella alpina 1S-4 is an industrial strain. In order to prepare host strains for a transformation system for this fungus, six uracil auxotrophs were obtained by means of random mutation with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). When the activities of orotate phosphoribosyl transferase (OPRTase, EC 2.4.2.10) and orotidine-5'-phosphate decarboxylase (OMPdecase, EC 4.1.1.23) were examined in the mutants and wild strain, OPRTase activity was found to be completely absent in all mutants, on the other hand, OMPdecase activity was intact. The genomic DNA and cDNA of the ura5 gene encoding OPRTase and the ura3 gene encoding OMPdecase were cloned and sequenced. The Ura5p deduced amino acid sequence of this fungus showed highest similarity to that of Vibrio cholerae classed among prokaryote. Furthermore, the mutational points in the ura5 genes of two selected mutants were identified; a base-replacement and a base-insertion.
(キーワード): Amino Acid Sequence / Base Sequence / Blotting, Southern / Cloning, Molecular / Culture Media / DNA, Complementary / DNA, Fungal / Gene Library / Genes, Fungal / Molecular Sequence Data / Mortierella / Mutation / Oils / Orotate Phosphoribosyltransferase / Orotidine-5'-Phosphate Decarboxylase / Uracil
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.68.277
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 14981288; ● Search Scopus @ Elsevier (PMID): 14981288; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.68.277

(DOI: 10.1271/bbb.68.277, PubMed: 14981288) Masataka Kajikawa, T. Katsuyuki Yamato, Yoshito Kohzu, Masutoshi Nojiri, Eiji Sakuradani, Sakayu Shimizu, Yasuyoshi Sakai, Hideya Fukuzawa and Kanji Ohyama :
Isolation and characterization of delta(6)-desaturase, an ELO-like enzyme and delta(5)-desaturase from the liverwort Marchantia polymorpha and production of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris.,
Plant Molecular Biology, Vol.54, No.3, 335-352, 2004.

(要約): The liverwort Marchantia polymorpha contains high proportions of arachidonic and eicosapentaenoic acids. In general, these C20 polyunsaturated fatty acids (PUFA) are synthesized from linoleic and alpha -linolenic acids, respectively, by a series of reactions catalyzed by Delta(6)-desaturase, an ELO-like enzyme involved in Delta(6) elongation and Delta(5)-desaturase. Here we report the isolation and characterization of the cDNAs, MpDES6, MpELO1 and MpDES5, coding for the respective enzymes from M. polymorpha. Co-expression of the MpDES6, MpELO1 and MpDES5 cDNAs resulted in the accumulation of arachidonic and eicosapentaenoic acids in the methylotrophic yeast Pichia pastoris. Interestingly, Delta(6) desaturation by the expression of the MpDES6 cDNA appears to occur both in glycerolipids and the acyl-CoA pool, although other lower-plant Delta(6)-desaturases are known to have a strong preference for glycerolipids.
(キーワード): Acetyl Coenzyme A / Amino Acid Sequence / Arachidonic Acid / Eicosapentaenoic Acid / Fatty Acid Desaturases / Fatty Acids, Unsaturated / Gene Expression Regulation, Enzymologic / Hepatophyta / Linoleoyl-CoA Desaturase / Mass Spectrometry / Molecular Sequence Data / Phylogeny / Pichia / Recombinant Proteins / Sequence Homology, Amino Acid / Substrate Specificity / alpha-Linolenic Acid
(出版サイトへのリンク): ● Publication site (DOI): 10.1023/B:PLAN.0000036366.57794.ee
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 15284491; ● Search Scopus @ Elsevier (PMID): 15284491; ● Search Scopus @ Elsevier (DOI): 10.1023/B:PLAN.0000036366.57794.ee

(DOI: 10.1023/B:PLAN.0000036366.57794.ee, PubMed: 15284491) Michihiko Kataoka, G A-R Delacruz-Hidalgo, Ali Muhammad Akond, Eiji Sakuradani, Keiko Kita and Sakayu Shimizu :
Gene cloning and overexpression of two conjugated polyketone reductases, novel aldo-keto reductase family enzymes, of Candida parapsilosis.,
Applied Microbiology and Biotechnology, Vol.64, No.3, 359-366, 2003.

(要約): The genes encoding two conjugated polyketone reductases (CPR-C1, CPR-C2) of Candida parapsilosis IFO 0708 were cloned and sequenced. The genes encoded a total of 304 and 307 amino acid residues for CPR-C1 and CPR-C2, respectively. The deduced amino acid sequences of the two enzymes showed high similarity to each other and to several proteins of the aldo-keto reductase (AKR) superfamily. However, several amino acid residues in putative active sites of AKRs were not conserved in CPR-C1 and CPR-C2. The two CPR genes were overexpressed in Escherichia coli. The E. coli transformant bearing the CPR-C2 gene almost stoichiometrically reduced 30 mg ketopantoyl lactone/ml to D-pantoyl lactone.
(キーワード): 4-Butyrolactone / Alcohol Oxidoreductases / Amino Acid Sequence / Base Sequence / Candida / Cloning, Molecular / Conserved Sequence / DNA, Fungal / Enzyme Stability / Escherichia coli / Gene Expression / Genes, Fungal / Hydrogen-Ion Concentration / Molecular Sequence Data / Recombinant Proteins / Sequence Analysis, DNA / Sequence Analysis, Protein / Sequence Homology, Amino Acid / Transformation, Bacterial
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-003-1484-3
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 14593510; ● Search Scopus @ Elsevier (PMID): 14593510; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-003-1484-3

(DOI: 10.1007/s00253-003-1484-3, PubMed: 14593510) Tetsuya Kurata, Chie Kawabata-Awai, Eiji Sakuradani, Sakayu Shimizu, Kiyotaka Okada and Takuji Wada :
The YORE-YORE gene regulates multiple aspects of epidermal cell differentiation in Arabidopsis.,
The Plant Journal : for Cell and Molecular Biology, Vol.36, No.1, 55-66, 2003.

(要約): We have identified a new Arabidopsis mutant, yore-yore (yre), which has small trichomes and glossy stems. Adhesion between epidermal cells was observed in the organs of the yre shoot. The cloned YRE had high homology to plant genes involved in epicuticular wax synthesis, such as ECERIFERUM1 (CER1) and maize GLOSSY1. The phenotype of transgenic plants harboring double-stranded RNA interference (dsRNAi) YRE was quite similar to that of the yre mutant. The amount of epicuticular wax extracted from leaves and stems of yre-1 was approximately one-sixth of that from the wild type. YRE promoter::GUS and in situ hybridization revealed that YRE was specifically expressed in cells of the L1 layer of the shoot apical meristem and young leaves, stems, siliques, and lateral root primordia. Strong expression was detected in developing trichomes. The trichome structure of cer1 was normal, whereas that of the yre cer1 double mutant was heavily deformed, indicating that epicuticular wax is required for normal growth of trichomes. Double mutants of yre and trichome-morphology mutants, glabra2 (gl2) and transparent testa glabra1 (ttg1), showed that the phenotype of the trichome structure was additive, suggesting that the wax-requiring pathway is distinct from the trichome development pathway controlled by GL2 and TTG1.
(キーワード): Amino Acid Sequence / Arabidopsis / Arabidopsis Proteins / Cell Differentiation / Cell Surface Extensions / Gene Expression Regulation, Developmental / Gene Expression Regulation, Plant / Molecular Sequence Data / Mutation / Phenotype / Plant Epidermis / Plants, Genetically Modified / RNA Interference / Sequence Homology, Amino Acid / Waxes
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12974811; ● Search Scopus @ Elsevier (PMID): 12974811

(PubMed: 12974811) Masataka Kajikawa, T. Katsuyuki Yamato, Hiroyuki Kanamaru, Eiji Sakuradani, Sakayu Shimizu, Hideya Fukuzawa, Yasuyoshi Sakai and Kanji Ohyama :
MpFAE3, a -Ketoacyl-CoA Synthase Gene in the Liverwort Marchantia polymorpha L., Is Preferentially Involved in Elongation of Palmitic Acid to Stearic Acid.,
Bioscience, Biotechnology, and Biochemistry, Vol.67, No.8, 1667-1674, 2003.

(要約): Fatty acid chain elongation is a crucial step in the biosynthesis of long chain fatty acids. An essential reaction in the elongation process is condensation of malonyl-CoA with acyl-CoA, which is catalyzed by beta-ketoacyl-CoA synthase (KCS) in plants. We have isolated and characterized the MpFAE3 gene, one of the KCS gene family in the liverwort Marchantia polymorpha. Transgenic M. polymorpha plants overexpressing MpFAE3 accumulate fatty acids 18:0, 20:0, and 22:0. In these plants, the amount of 16:0 is reduced to 50% of wild type. In a heterologous assay, transgenic methylotrophic yeast expressing the MpFAE3 gene accumulates fatty acid 18:0 and generates several longer fatty acids which are not detectable in the control, accompanied by a decrease of 16:0. These observations indicate that the MpFAE3 protein is preferentially involved in the elongation of 16:0 to 18:0 and also in the subsequent steps of 18:0 to 20:0 and 20:0 to 22:0 in M. polymorpha.
(キーワード): 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Amino Acid Sequence / Blotting, Northern / Blotting, Southern / Fatty Acids / 遺伝子発現 (gene expression) / Genes, Plant / Hepatophyta / Molecular Sequence Data / Palmitic Acid / Phylogeny / Plants, Genetically Modified / Sequence Alignment / Stearic Acids
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.67.1667
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12951498; ● Summary page in Scopus @ Elsevier: 2-s2.0-2442466138

(DOI: 10.1271/bbb.67.1667, PubMed: 12951498, Elsevier: Scopus) Eiji Sakuradani and Sakayu Shimizu :
Gene Cloning and Functional Analysis of a Second 6-Fatty Acid Desaturase from an Arachidonic Acid-producing Mortierella Fungus,
Bioscience, Biotechnology, and Biochemistry, Vol.67, No.4, 704-711, 2003.

(要約): We demonstrated that Mortierella alpina 1S-4 has two delta 6-desaturases, which are involved in the desaturation of linoleic acid to gamma-linolenic acid. For one of the two delta 6-desaturases, designated as delta 6I, gene cloning and its heterologous expression in a fungus, Aspergillus oryzae, has previously been reported. In addition, we indicated in this paper that there is an isozyme of the two delta 6-desaturases, designated as delta 6II, in M. alpina 1S-4. The predicted amino acid sequences of the Mortierella delta 6-desaturases were similar to those of ones from other organisms, i.e. borage and Caenorhabditis elegans, and had a cytochrome b5-like domain at the N-terminus, being different from the yeast delta 9-desaturase, which has the corresponding domain at the C-terminus. The full-length delta 6II cDNA was expressed in A. oryzae, resulting in the accumulation of gamma-linolenic acid (which was not detected in the control Aspergillus) up to 37% of the total fatty acids. The analysis of real-time quantitative PCR (RTQ-PCR) showed that the quantity of delta 6I RNA was 2.4-, 9-, and 17-fold higher than that of delta 6II RNA on 2, 3, and 4 days in M. alpina 1S-4, respectively. M. alpina 1S-4 is the first fungus to be confirmed to have two functional delta 6-desaturase genes.
(キーワード): Amino Acid Sequence / Arachidonic Acid / Aspergillus oryzae / Cloning, Molecular / Fatty Acid Desaturases / Genes, Fungal / Isoenzymes / Linoleoyl-CoA Desaturase / Molecular Sequence Data / Mortierella / RNA
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.67.704
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12784608; ● Search Scopus @ Elsevier (PMID): 12784608; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.67.704

(DOI: 10.1271/bbb.67.704, PubMed: 12784608) Masataka Kajikawa, Shohei Yamaoka, T. Katsuyuki Yamato, Hiroyuki Kanamaru, Eiji Sakuradani, Sakayu Shimizu, Hideya Fukuzawa and Kanji Ohyama :
Functional analysis of a beta-ketoacyl-CoA synthase gene, MpFAE2, by gene silencing in the liverwort Marchantia polymorpha L.,
Bioscience, Biotechnology, and Biochemistry, Vol.67, No.3, 605-612, 2003.

(要約): We have isolated a beta-ketoacyl CoA synthase (KCS) gene, MpFAE2, from a liverwort, Marchantia polymorpha, and identified its substrate specificity using the technique of dsRNA-mediated gene silencing and overexpression. KCS catalyzes an essential reaction in the fatty acid elongation process, i.e., condensation of malonyl-CoA with acyl-CoA. By introducing a construct with a hairpin structure containing a partial MpFAE2 gene, the level of the MpFAE2 gene expression was suppressed constitutively. The transgenic plants showed a specific accumulation of fatty acid 18:0. In contrast, in transgenic M. polymorpha plants overexpressing the MpFAE2 gene, fatty acid 22:0 is accumulated. These results indicate that the MpFAE2 gene product catalyzes the elongation steps of 18:0 to 20:0 and possibly also of 20:0 to 22:0.
(キーワード): 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Acetyl Coenzyme A / Amino Acid Sequence / Blotting, Northern / Blotting, Southern / Fatty Acids / Gene Silencing / Hepatophyta / Malonyl Coenzyme A / Molecular Sequence Data / Nucleic Acid Conformation / Plants, Genetically Modified / RNA Interference / Sequence Alignment / Sequence Homology, Amino Acid / Substrate Specificity
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.67.605
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12723610; ● Search Scopus @ Elsevier (PMID): 12723610; ● Search Scopus @ Elsevier (DOI): 10.1271/bbb.67.605

(DOI: 10.1271/bbb.67.605, PubMed: 12723610) Kohsuke Honda, Michihiko Kataoka, Eiji Sakuradani and Sakayu Shimizu :
Role of Acinetobacter calcoaceticus 3,4-dihydrocoumarin hydrolase in oxidative stress defence against peroxoacids.,
European Journal of Biochemistry, Vol.270, No.3, 486-494, 2003.

(要約): The physiological role of a bifunctional enzyme, 3,4-dihydrocoumarin hydrolase (DCH), which is capable of both hydrolysis of ester bonds and organic acid-assisted bromination of organic compounds, was investigated. Purified DCH from Acinetobacter calcoaceticus F46 catalysed dose- and time-dependent degradation of peracetic acid. The gene (dch) was cloned from the chromosomal DNA of the bacterium. The dch ORF was 831 bp long, corresponding to a protein of 272 amino acid residues, and the deduced amino acid sequence showed high similarity to those of bacterial serine esterases and perhydrolases. The dch gene was disrupted by homologous recombination on the A. calcoaceticus genome. The dch disruptant strain was more sensitive to growth inhibition by peracetic acid than the parent strain. On the other hand, the recombinant Escherichia coli cells expressing dch were more resistant to peracetic acid. A putative catalase gene was found immediately downstream of dch, and Northern blot hybridization analysis revealed that they are transcribed as part of a polycistronic mRNA. These results suggested that in vivo DCH detoxifies peroxoacids in conjunction with the catalase, i.e. peroxoacids are first hydrolysed to the corresponding acids and hydrogen peroxide by DCH, and then the resulting hydrogen peroxide is degraded by the catalase.
(キーワード): Acinetobacter calcoaceticus / Amino Acid Sequence / Blotting, Northern / DNA Primers / DNA, Bacterial / Gene Expression Regulation, Bacterial / Hydrolases / Hydrolysis / Molecular Sequence Data / Oxidants / Oxidative Stress / Peracetic Acid / Peroxides / Polymerase Chain Reaction / Recombinant Proteins / Recombination, Genetic / Sequence Homology, Amino Acid / Transformation, Bacterial
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12542698; ● Search Scopus @ Elsevier (PMID): 12542698

(PubMed: 12542698) Eiji Sakuradani, Nozomu Kamada, Yuriko Hirano, Mitsuhiro Nishihara, Hiroshi Kawashima, Kengo Akimoto, Kenichi Higashiyama, Jun Ogawa and Sakayu Shimizu :
Production of 5,8,11-eicosatrienoic acid by a delta5 and delta6 desaturation activity-enhanced mutant derived from a delta12 desaturation activity-defective mutant of Mortierella alpina 1S-4.,
Applied Microbiology and Biotechnology, Vol.60, No.3, 281-287, 2002.

(要約): Enhanced production of 5,8,11-eicosatrienoic acid (Mead acid, 20:3omega9) was attained with a mutant fungus, Mortierella alpina JT-180, derived from delta12 desaturation activity-defective and delta6 desaturation activity-enhanced M. alpina M209-7. Production of 20:3omega9 by JT-180 was 1.4 times greater than that of the parent strain M209-7. This is thought to be due to its enhanced Delta5 desaturation activity, which was 3.3 times higher than that of M209-7. In both strains, 78.5-80.4% of the total lipids comprised triacylglycerol (TG), and 76.6-79.0% of 20:3omega9 was present in TG. Comparing the fatty acid compositions among various lipid species, the highest percentages (24.1-37.6%) of 20:3omega9 in total lipids were found in phosphatidylcholine. For optimization of 20:3omega9 production by JT-180, a glucose concentration of 4% in the culture medium and shifting of the growth temperature from 28 degrees C to 20 degrees C on the 2nd day were shown to be effective. Under optimal conditions, 20:3omega9 production by JT-180 reached 1.92 g/l culture medium in a 10-l jar fermentor (corresponding to 81.5 mg/g dry mycelia and 18.3% of total fatty acids), which is greater than that reported previously from M209-7 (1.65 g/l).
(キーワード): 8,11,14-Eicosatrienoic Acid / Culture Media / Fatty Acid Desaturases / Fatty Acids / Glucose / Mortierella / Mutation / Temperature
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-002-1128-z
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 12436308; ● Search Scopus @ Elsevier (PMID): 12436308; ● Search Scopus @ Elsevier (DOI): 10.1007/s00253-002-1128-z

(DOI: 10.1007/s00253-002-1128-z, PubMed: 12436308) Saeree Jareonkitmongkol, Eiji Sakuradani and Sakayu Shimizu :
Isolation and characterization of a Δ9-desaturation-defective mutant of an arachidonic acid-producing fungus, Mortierella alpina 1S-4,
Journal of the American Oil Chemists' Society, Vol.79, No.10, 1021-1026, 2002.

(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s11746-002-0596-1
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1007/s11746-002-0596-1

(DOI: 10.1007/s11746-002-0596-1) Chee-Leong Soong, Jun Ogawa, Eiji Sakuradani and Sakayu Shimizu :
Barbiturase, a novel zinc-containing amidohydrolase involved in oxidative pyrimidine metabolism.,
The Journal of Biological Chemistry, Vol.277, No.9, 7051-7058, 2001.

(要約): Barbiturase, which catalyzes the reversible amidohydrolysis of barbituric acid to ureidomalonic acid in the second step of oxidative pyrimidine degradation, was purified to homogeneity from Rhodococcus erythropolis JCM 3132. The characteristics and gene organization of barbiturase suggested that it is a novel zinc-containing amidohydrolase that should be grouped into a new family of the amidohydrolases superfamily. The amino acid sequence of barbiturase exhibited 48% identity with that of herbicide atrazine-decomposing cyanuric acid amidohydrolase but exhibited no significant homology to other proteins, indicating that cyanuric acid amidohydrolase may have evolved from barbiturase. A putative uracil phosphoribosyltransferase gene was found upstream of the barbiturase gene, suggesting mutual interaction between pyrimidine biosynthesis and oxidative degradation. Metal analysis with an inductively coupled radiofrequency plasma spectrophotometer revealed that barbiturase contains approximately 4.4 mol of zinc per mol of enzyme. The homotetrameric enzyme had K(m) and V(max) values of 1.0 mm and 2.5 micromol/min/mg of protein, respectively, for barbituric acid. The enzyme specifically acted on barbituric acid, and dihydro-l-orotate, alloxan, and cyanuric acid competitively inhibited its activity. The full-length gene encoding the barbiturase (bar) was cloned and overexpressed in Escherichia coli. The kinetic parameters and physicochemical properties of the cloned enzyme were apparently similar to those of the wild-type.
(キーワード): Amidohydrolases / Amino Acid Motifs / Amino Acid Sequence / Aminohydrolases / Base Sequence / Binding Sites / Binding, Competitive / Cloning, Molecular / Electrophoresis, Polyacrylamide Gel / Escherichia coli / Hydrogen-Ion Concentration / Kinetics / Malonates / Models, Chemical / Molecular Sequence Data / Oxygen / Peptides / Phylogeny / Pyrimidines / Recombinant Proteins / Rhodococcus / Sequence Homology, Amino Acid / Spectrophotometry / Temperature / Triazines / Zinc
(出版サイトへのリンク): ● Publication site (DOI): 10.1074/jbc.M110784200
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 11748240; ● Search Scopus @ Elsevier (PMID): 11748240; ● Search Scopus @ Elsevier (DOI): 10.1074/jbc.M110784200

(DOI: 10.1074/jbc.M110784200, PubMed: 11748240) Milan Certik, Eiji Sakuradani, Michihiko Kobayashi and Sakayu Shimizu :
Characterization of the Second Form of NADH-Cytochrome b_5 Reductase Gene from Arachidonic Acid-Producing Fungus Mortierella alpina 1S-4,
Journal of Bioscience and Bioengineering, Vol.88, No.6, 667-671, 1999.

(要約): The second type of cytochrome b_5 reductase (Cb5R-II) gene was characterized in the arachidonic acidproducing fungus Mortierella alpina 1S-4. Its cDNA (897 bp) and predicted amino acid (298 aa) sequences show more than 70% similarity to the previously isolated first type of Cb5R. Highly conserved exon-intron organization suggests that the two genes evolved from the duplication of a common ancestral gene. Cb5R-II has a flavin-binding domain at its highly hydrophobic N-terminal and an NADH-binding domain at the C-terminal. In comparison with Cb5R genes from other sources, high homology (46-54%) was found for yeast and plant genes. Phylogenetic analysis revealed that microbial and plant Cb5R genes represent a gene family evolved from one prototype and are different from mammalian Cb5R genes.
(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1523669555486568832

(CiNii: 1523669555486568832) Nozomu Kamada, Hiroshi Kawashima, Eiji Sakuradani, Kengo Akimoto, Jun Ogawa and Sakayu Shimizu :
Production of 8,11-cis-eicosadienoic acid by a Δ5 and Δ12 desaturase-defective mutant derived from the arachidonic acid-producing fungus Mortierella alpina 1S-4,
Journal of the American Oil Chemists' Society, Vol.76, No.11, 1269-1274, 1999. Eiji Sakuradani, Michihiko Kobayashi and Sakayu Shimizu :
Delta6-fatty acid desaturase from an arachidonic acid-producing Mortierella fungus. Gene cloning and its heterologous expression in a fungus, Aspergillus.,
Gene, Vol.238, No.2, 445-453, 1999.

(要約): A DNA fragment was cloned from the fungal strain, Mortierella alpina 1S-4 (which is used industrially to produce arachidonic acid), after PCR amplification with oligonucleotide primers designed based on the sequence information for delta6-desaturase genes (from borage and Caenorhabditis elegans), which are involved in the desaturation of linoleic acid (delta9, delta12-18:2) to gamma-linolenic acid (delta6, delta9, delta12-18:3). This fragment was used as a probe to isolate a cDNA clone with an open reading frame encoding 457 amino acids from a M. calpina 1S-4 library. The predicted amino-acid sequence showed similarity to those of the above delta6-desaturases, and contained a cytochrome b5-like domain at the N-terminus, being different from the yeast delta9-desaturase which has the corresponding domain at the C-terminus. The full-length cDNA clone was expressed under the control of the amyB promoter in a filamentous fungus, Aspergillus oryzae, resulting in the accumulation of gamma-linolenic acid (which was not detected in the control Aspergillus) to the level of 25.2% of the total fatty acids. These findings revealed that the recombinant product has delta6-desaturase activity. The Mortierella delta6-desaturase is the first to be reported in fungi.
(キーワード): Amino Acid Sequence / Arachidonic Acid / Aspergillus oryzae / Base Sequence / Blotting, Southern / Cloning, Molecular / DNA, Complementary / Fatty Acid Desaturases / Gas Chromatography-Mass Spectrometry / Linoleoyl-CoA Desaturase / Lipids / Molecular Sequence Data / Mortierella / Open Reading Frames / Sequence Homology, Amino Acid
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10570972; ● Search Scopus @ Elsevier (PMID): 10570972

(PubMed: 10570972) Eiji Sakuradani, Michihiko Kobayashi and Sakayu Shimizu :
Identification of an NADH-cytochrome b(5) reductase gene from an arachidonic acid-producing fungus, Mortierella alpina 1S-4, by sequencing of the encoding cDNA and heterologous expression in a fungus, Aspergillus oryzae.,
Applied and Environmental Microbiology, Vol.65, No.9, 3873-3879, 1999.

(要約): Based on the sequence information for bovine and yeast NADH-cytochrome b(5) reductases (CbRs), a DNA fragment was cloned from Mortierella alpina 1S-4 after PCR amplification. This fragment was used as a probe to isolate a cDNA clone with an open reading frame encoding 298 amino acid residues which show marked sequence similarity to CbRs from other sources, such as yeast (Saccharomyces cerevisiae), bovine, human, and rat CbRs. These results suggested that this cDNA is a CbR gene. The results of a structural comparison of the flavin-binding beta-barrel domains of CbRs from various species and that of the M. alpina enzyme suggested that the overall barrel-folding patterns are similar to each other and that a specific arrangement of three highly conserved amino acid residues (i.e., arginine, tyrosine, and serine) plays a role in binding with the flavin (another prosthetic group) through hydrogen bonds. The corresponding genomic gene, which was also cloned from M. alpina 1S-4 by means of a hybridization method with the above probe, had four introns of different sizes. These introns had GT at the 5' end and AG at the 3' end, according to a general GT-AG rule. The expression of the full-length cDNA in a filamentous fungus, Aspergillus oryzae, resulted in an increase (4.7 times) in ferricyanide reduction activity involving the use of NADH as an electron donor in the microsomes. The M. alpina CbR was purified by solubilization of microsomes with cholic acid sodium salt, followed by DEAE-Sephacel, Mono-Q HR 5/5, and AMP-Sepharose 4B affinity column chromatographies; there was a 645-fold increase in the NADH-ferricyanide reductase specific activity. The purified CbR preferred NADH over NADPH as an electron donor. This is the first report of an analysis of this enzyme in filamentous fungi.
(キーワード): Amino Acid Sequence / Animals / Arachidonic Acid / Aspergillus oryzae / Cattle / Cloning, Molecular / Cytochrome Reductases / Cytochrome-B(5) Reductase / DNA, Complementary / Humans / Molecular Sequence Data / Mortierella / Rats / Sequence Alignment / Sequence Analysis, DNA
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10473389; ● Search Scopus @ Elsevier (PMID): 10473389

(PubMed: 10473389) Eiji Sakuradani, Michihiko Kobayashi, Toshihiko Ashikari and Sakayu Shimizu :
Identification of Delta12-fatty acid desaturase from arachidonic acid-producing mortierella fungus by heterologous expression in the yeast Saccharomyces cerevisiae and the fungus Aspergillus oryzae.,
European Journal of Biochemistry, Vol.261, No.3, 812-820, 1999.

(要約): Based on the sequence information for the omega3-desaturase genes (from Brassica napus and Caenorhabditis elegans), which are involved in the desaturation of linoleic acid (Delta9, Delta12-18 : 2) to alpha-linolenic acid (Delta9, Delta12, Delta15-18 : 3), a cDNA was cloned from the filamentous fungal strain, Mortierella alpina 1S-4, which is used industrially to produce arachidonic acid. Homology analysis with protein databases revealed that the amino acid sequence showed 43.7% identity as the highest match with the microsomal omega6-desaturase (from Glycine max, soybean), whereas it exhibited 38.9% identity with the microsomal omega3-desaturase (from soybean). The evolutionary implications of these enzymes will be discussed. The cloned cDNA was confirmed to encode a Delta12-desaturase, which was involved in the desaturation of oleic acid (Delta9-18 : 1) to linoleic acid, by its expression in both the yeast Saccharomyces cerevisiae and the fungus Aspergillus oryzae. Analysis of the fatty acid composition of yeast and fungus transformants demonstrated that linoleic acid (which was not contained in the control strain of S. cerevisiae) was accumulated in the yeast transformant and that the fungal transformant contained a large amount of linoleic acid (71.9%). Genomic Southern blot analysis of the transformants with the Mortierella Delta12-desaturase gene as a probe confirmed integration of this gene into the genome of A. oryzae. The M. alpina 1S-4 Delta12-desaturase is the first example of a cloned nonplant Delta12-desaturase.
(キーワード): Amino Acid Sequence / Arachidonic Acid / Aspergillus oryzae / Base Sequence / Blotting, Southern / Cloning, Molecular / DNA Primers / DNA, Complementary / Fatty Acid Desaturases / Molecular Sequence Data / Mortierella / Saccharomyces cerevisiae / Sequence Homology, Amino Acid
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10215899; ● Search Scopus @ Elsevier (PMID): 10215899

(PubMed: 10215899) Eiji Sakuradani, Michihiko Kobayashi and Sakayu Shimizu :
Delta 9-fatty acid desaturase from arachidonic acid-producing fungus. Unique gene sequence and its heterologous expression in a fungus, Aspergillus.,
European Journal of Biochemistry, Vol.260, No.1, 208-216, 1999.

(要約): Based on the sequence information for delta 9-desaturase genes (from rat, mouse and yeast), which are involved in the desaturation of palmitic acid and stearic acid to palmitoleic acid and oleic acid, respectively, the corresponding cDNA and genomic gene were cloned from the fungal strain, Mortierella alpina 1S-4, which industrially produces arachidonic acid. There was a cytochrome b5-like domain linked to the carboxyl terminus of this Mortierella desaturase, as also seen in the yeast delta 9-desaturase. The Mortierella delta 9-desaturase genomic gene had only one intron, in which a novel phenomenon was observed: there was a GC-end at the 5'-terminus instead of a GT-end that is, in general, found in introns of eukaryotic genes. The full-length cDNA clone was expressed under the control of an amyB promoter in a filamentous fungus, Aspergillus oryzae, resulting in drastic changes in the fatty acid composition in the transformant cells; the contents of palmitoleic acid (16:1) and oleic acid (18:1) increased significantly, with accompanying decreases in palmitic acid (16:0) and stearic acid (18:0). These changes were controlled by the addition of maltose as a carbon source to the medium. Also, the expression of the gene caused a significant change in the lipid composition in the Aspergillus transformant. Genomic Southern blot analysis of the transformant with the Mortierella delta 9-desaturase gene as a probe confirmed the integration of this gene into the genome of A. oryzae.
(キーワード): Amino Acid Sequence / Arachidonic Acid / Aspergillus oryzae / Base Sequence / Blotting, Southern / Cloning, Molecular / Fatty Acid Desaturases / Fatty Acids / Fungal Proteins / Gene Expression Regulation, Enzymologic / Gene Expression Regulation, Fungal / Lipids / Molecular Sequence Data / Mortierella / Sequence Analysis, DNA / Sequence Homology, Amino Acid
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 10091601; ● Search Scopus @ Elsevier (PMID): 10091601

(PubMed: 10091601) Michihiko Kobayashi, Eiji Sakuradani and Sakayu Shimizu :
Genetic Analysis of Cytochrome b5 from Arachidonic Acid-Producing Fungus, Mortierella alpina 1S-4: Cloning, RNA Editing and Expression of the Gene in Escherichia coli, and Purification and Characterization ofthe Gene Product.,
The Journal of Biochemistry, Vol.125, No.6, 1094-1103, 1999.

(要約): Information on the amino acid sequences of the internal peptide fragments of cytochrome b5 from Mortierella hygrophila was used to prepare synthetic oligonucleotides as primers for the polymerase chain reaction. A 100-base DNA fragment was thus amplified, by using a genomic gene from Mortierella alpina 1S-4 as a template, which produced polyunsaturated fatty acids such as arachidonic acid. The amplified DNA fragment was used as the probe to clone both a 523-base cDNA fragment and a 2.1-kilobase Sal1-NruI genomic fragment coding for the whole M. alpina 1S-4 cytochrome b5. On the basis of nucleotide sequences of both cytochrome b5 genomic gene and cDNA, the genomic cytochrome b5 gene was found to consist of four exons and three introns. A novel type of RNA editing, in which the cDNA included either guanine insertion or adenineguanine substitution at one base upstream of poly(A), was interestingly observed. The deduced amino acid sequence of M. alpina 1S-4 cytochrome b5 showed significant similarities with those of cytochrome b5s from other organisms such as rat, chicken, and yeast. The soluble form of the cytochrome b5 gene was expressed to 16% of the total soluble protein in Eseherichia coli. The holo-cytochrome b5 accounted for 8% of the total cytochrome b5 in the transformants. The purified cytochrome b5 showed the oxidized and reduced absorbance spectra characteristic of fungal microsomal cytochrome b5.
(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1574231877859637376

(CiNii: 1574231877859637376) Hiroshi Kawashima, Eiji Sakuradani, Nozomu Komada, Kengo Akimoto, Kyoko Konishi, Jun Ogawa and Sakayu Shimizu :
Production of 8,11,14,17-cis-eicosatetraenoic acid (20:4ω3) by a Δ5 and Δ12 desaturase- defective mutant of an arachidonic acid-producing fungus Mortierella alpina 1S-4,
Journal of the American Oil Chemists' Society, Vol.75, No.11, 1495-1500, 1998. Hiroshi Kawashima, Nozomu Kamada, Eiji Sakuradani, Saeree Jareonkitmongkol, Kengo Akimoto and Sakayu Shimizu :
Production of 8,11,14,17-cis-eicosatetraenoic acid by 5 desaturase-defective mutants of an arachidonic acid-pruducing fungus, Mortierella alpina.,
Journal of the American Oil Chemists' Society, Vol.74, No.4, 455-459, 1997.

(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1572824500672907904

(CiNii: 1572824500672907904) Saeree Jareonkitmongkol, Eiji Sakuradani and Sakayu Shimizu :
Isolation and characterization of an ω3- desaturation-defective mutant of an arachidonic acid-producing fungus, Mortierella alpina 1S-4,
Arch. Microbiol., Vol.161, No.4, 316-319, 1994. Saeree Jareonkitmongkol, Eiji Sakuradani and Sakayu Shimizu :
A Novel Delta5-Desaturase-Defective Mutant of Mortierella alpina 1S-4 and Its Dihomo-gamma-Linolenic Acid Productivity.,
Applied and Environmental Microbiology, Vol.59, No.12, 4300-4304, 1993.

(要約): A novel Delta5-desaturase-defective mutant was derived from an arachidonic acid-producing fungus, Mortierella alpina 1S-4, after treating the parental spores with N-methyl-N'-nitro-N-nitrosoguanidine. The mutant produced only a trace (about 1%) amount of arachidonic acid, and the ratio of dihomo-gamma-linolenic acid (DGLA) to total fatty acids in each lipid class was markedly high, accounting for as much as 60% in phosphatidylcholine. Under submerged batch culture conditions, the mutant produced 2.4 g of DGLA per liter (43.3% of total fatty acids) when grown at 28 degrees C for 7 days in a 5-liter jar fermentor. The other major (more that 1%) fatty acids were palmitic acid (21.2%), stearic acid (9.6%), oleic acid (14.3%), linoleic acid (4.4%), and gamma-linolenic acid (5.8%). About 80 mol% of the DGLA produced was found in triacylglycerol.
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 16349126; ● Search Scopus @ Elsevier (PMID): 16349126

(PubMed: 16349126)

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Hiroshi Kikukawa, Kenshi Watanabe, Shigenobu Kishino, Michiki Takeuchi, Akinori Ando, Yoshihiro Izumi and Eiji Sakuradani :
Recent trends in the field of lipid engineering,
Journal of Bioscience and Bioengineering, Vol.133, No.5, 405-413, May 2022.

(要約): Lipid engineering related to biological functions has made remarkable progress in the fields of microbial production of functional lipids, metabolic engineering of microorganisms, elucidation of physiological functions of rare lipids, lipid-related enzyme engineering, and lipid analysis techniques. Various rare lipids are produced by utilizing microorganisms and their enzymes. It is also becoming clear that the rare lipids produced by intestinal bacteria contribute significantly to human health. Technological advances related to identification of lipid structures and quantification of lipids have led to such discoveries in the field of lipid engineering. This article reviews the latest findings that are attracting attention in the field of lipid engineering related to biological functions.
(キーワード): Humans / Lipids / Metabolic Engineering
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2022.02.001
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 35249828; ● Search Scopus @ Elsevier (PMID): 35249828; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiosc.2022.02.001

(DOI: 10.1016/j.jbiosc.2022.02.001, PubMed: 35249828) 安藤晃規, 奥田知生, 菊川寛史, 阪本鷹行, 櫻谷英治, 小川順 :
油糧微生物による種々のω3脂肪酸含有油脂の発酵生産,
月刊バイオインダストリー, Vol.37, No.10, 3-13, 2020年10月. 小川順, 櫻谷英治, 岸野重信, 安藤晃規, 清水昌 :
高度不飽和脂肪酸・共役脂肪酸含有油脂の微生物生産,
微生物を活用した新世代の有用物質生産技術《普及版》, 159-164, 2019年8月. Hiroshi Kikukawa, Eiji Sakuradani, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Arachidonic acid production by the oleaginous fungus Mortierella alpina 1S-4: A review,
Journal of Advanced Research, Vol.11, 15-22, Feb. 2018.

(要約): The filamentous fungus Mortierella alpina 1S-4 is capable of accumulating a large amount of triacylglycerol containing C20 polyunsaturated fatty acids (PUFAs). Indeed, triacylglycerol production by M. alpina 1S-4 can reach 20 g/L of culture broth, and the critical cellular signaling and structural PUFA arachidonic acid (ARA) comprises 30% 70% of the total fatty acid. The demonstrated health benefits of functional PUFAs have in turn encouraged the search for rich sources of these compounds, including fungal strains showing enhanced production of specific PUFAs. Screening for mutants and targeted gene manipulation of M. alpina 1S-4 have elucidated the functions of various enzymes involved in PUFA biosynthesis and established lines with improved PUFA productivity. In some cases, these strains have been used for indistrial-scale production of PUFAs, including ARA. In this review, we described practical ARA production through mutant breeding, functional analyses of genes encoding enzymes involved in PUFA biosynthesis, and recent advances in the production of specific PUFAs through molecular breeding of M. alpina 1S-4.
(徳島大学機関リポジトリ): ● Metadata: 115482
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jare.2018.02.003
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.1016/j.jare.2018.02.003

(徳島大学機関リポジトリ: 115482, DOI: 10.1016/j.jare.2018.02.003) Eiji Sakuradani, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Metabolic engineering for the production of polyunsaturated fatty acids by oleaginous fungus Mortierella alpina 1S-4,
Journal of Bioscience and Bioengineering, Vol.116, No.4, 417-422, Oct. 2013.

(要約): Researches related with the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have been conducted in various fields with a view to health and dietary requirements. Novel rich sources other than known natural sources such as plant seeds and fish oils are required for increasing demands of PUFAs. The filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, i.e., ones reaching 20 g/l in concentration and containing 30-70% arachidonic acid as total fatty acids. Various mutants derived from M. alpina 1S-4 have led to the production of oils containing various PUFAs. Molecular breeding of M. alpina strains by means of manipulation of the genes involved in PUFA biosynthesis facilitates improvement of PUFA productivity and elucidation of the functions of their enzymes. This review describes practical PUFA production through mutant breeding, functional analyses of the genes of the enzymes involved in PUFA biosynthesis, and recent advances in unique PUFA production through molecular breeding.
(キーワード): Mortierella alpina / Polyunsaturated fatty acid / Arachidonic acid (AA) / Eicosapentaenoic acid (EPA) / Molecular breeding / Fatty acid desaturase
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2013.04.008
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23648102; ● CiNii @ 国立情報学研究所 (CRID): 1050282810754539776; ● Summary page in Scopus @ Elsevier: 2-s2.0-84883284901

(DOI: 10.1016/j.jbiosc.2013.04.008, PubMed: 23648102, CiNii: 1050282810754539776, Elsevier: Scopus) 櫻谷英治, 岸野重信, 安藤晃規, 新井洋由, 植田和光, 小川順 :
オメガ3脂肪酸含有油脂の微生物による生産と変換,
月刊バイオインダストリー, Vol.30, No.2, 32-39, 2013年2月.

(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1523669555286561408

(CiNii: 1523669555286561408) Jun Ogawa, Eiji Sakuradani, Shigenobu Kishino, Akinori Ando, Kenzo Yokozeki and Sakayu Shimizu :
Polyunsaturated fatty acids production and transformation by Mortierella alpina and anaerobic bacteria,
European Journal of Lipid Science and Technology : EJLST, Vol.114, No.10, 1107-1113, Oct. 2012.

(出版サイトへのリンク): ● Publication site (DOI): 10.1002/ejlt.201200069
(文献検索サイトへのリンク): ● Summary page in Scopus @ Elsevier: 2-s2.0-84867673153

(DOI: 10.1002/ejlt.201200069, Elsevier: Scopus) 櫻谷英治, 安藤晃規, 清水昌, 小川順 :
アラキドン酸および関連高度不飽和脂肪酸を含む微生物油脂生産,
オレオサイエンス, Vol.12, No.7, 263-272, 2012年7月.

(要約): 高度不飽和脂肪酸(PUFA)のような機能性脂質の応用研究は，「健康」や「栄養」に関わる様々な分野で行われてきた．特に注目されてきた炭素鎖長20以上のPUFAの供給源は，鶏卵に含まれるアラキドン酸や魚油に含まれるn-3 PUFAが主なものであった．炭素鎖長20以上のPUFAを生産する微生物スクリーニングにより，アラキドン酸を豊富に含むトリアシルグリセロールを生産する油糧微生物Mortierella alpina 1S-4が見いだされた．M. alpina 1S-4において，油脂の生産量は20 g/lに達し，総脂肪酸に占めるアラキドン酸の割合は30∼70%を占める．これまでに，M. alpina 1S-4から様々な変異株が誘導され，多様なPUFAが生産されるようになった．さらに，M. alpina 1S-4の形質転換系の開発により，PUFA生合成に関わる酵素遺伝子の解明やPUFA生産の向上が可能となった．本稿では，M. alpina 1S-4の育種(変異株育種と分子育種を含む)によるアラキドン酸とその関連PUFAの実用的な微生物生産について報告する．
(キーワード): Mortierella alpina / polyunsaturated fatty acid / arachidonic acid / molecular breeding / fatty acid desaturase
(出版サイトへのリンク): ● Publication site (DOI): 10.5650/oleoscience.12.263
(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1390282679461348608; ● Search Scopus @ Elsevier (DOI): 10.5650/oleoscience.12.263

(DOI: 10.5650/oleoscience.12.263, CiNii: 1390282679461348608) 小川順, 櫻谷英治, 岸野重信, 日比慎, 安藤晃規 :
バイオプラ最前線-バイオマスからのモノマー生産を支援するバイオリピッドプラットフォームの構築-,
バイオプラジャーナル, Vol.11, No.4, 13-19, 2012年2月. 小川順, 櫻谷英治, 岸野重信, 日比慎, 堀之内伸行, 横関健三, 清水昌 :
代謝的視点からの機能探索に基づく産業プロセス用酵素開発,
月刊バイオインダストリー, Vol.28, No.9, 7-13, 2011年8月. 小川順, 櫻谷英治, 安藤晃規, 清水昌 :
第6回油脂生産性糸状菌Mortierella alpinaによる機能性脂質生産連載「微生物の産業利用-はたらく有用微生物-」,
Microbiol. Cult. Coll., Vol.27, No.6, 25-29, 2011年6月. Eiji Sakuradani :
Advances in the production of various polyunsaturated fatty acids through oleaginous fungus Mortierella alpina breeding,
Bioscience, Biotechnology, and Biochemistry, Vol.74, No.5, 908-917, May 2010.

(要約): Studies of the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have been conducted in various fields with a view to health and dietary requirements in a search for novel, rich sources. The filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, i.e., ones reaching 20 g/l in concentration and containing 30-70% arachidonic acid as total fatty acids. Various mutants derived from M. alpina 1S-4 have led to the production of oils containing various PUFAs. Molecular breeding of M. alpina strains by means of manipulation of the genes involved in PUFA biosynthesis facilitates improvement of PUFA productivity and elucidation of the functions of their enzymes. This review describes practical PUFA production through mutant breeding, functional analyses of the genes of the enzymes involved in PUFA biosynthesis, and recent advances in unique PUFA production through molecular breeding.
(キーワード): Acetyltransferases / Animals / Fatty Acid Desaturases / Fatty Acids, Unsaturated / Genetic Engineering / Humans / Mortierella / Mutation
(出版サイトへのリンク): ● Publication site (DOI): 10.1271/bbb.100001
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 20460719; ● Summary page in Scopus @ Elsevier: 2-s2.0-77955407890

(DOI: 10.1271/bbb.100001, PubMed: 20460719, Elsevier: Scopus) Eiji Sakuradani and Sakayu Shimizu :
Single cell oil production by Mortierella alpina,
Journal of Biotechnology, Vol.144, No.1, 31-36, Oct. 2009.

(要約): A filamentous fungus, Mortierella alpina 1S-4, was obtained, through extensive screening, as an potential producer of triacylglycerol containing C20 polyunsaturated fatty acids (PUFAs) such as arachidonic acid. With this discovery as a starting point, we conducted employing methods from metabolic engineering and molecular biology for controlling cultures and breeding mutant strains. These parental and mutant strains are now used for large-scale production of a variety of PUFAs.
(キーワード): Arachidonic Acid / Genetic Engineering / Mortierella / Mutation / Oils
(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiotec.2009.04.012
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19409938; ● Search Scopus @ Elsevier (PMID): 19409938; ● Search Scopus @ Elsevier (DOI): 10.1016/j.jbiotec.2009.04.012

(DOI: 10.1016/j.jbiotec.2009.04.012, PubMed: 19409938) Eiji Sakuradani, Akinori Ando, Jun Ogawa and Sakayu Shimizu :
Improved production of various polyunsaturated fatty acids through filamentous fungus Mortierella alpina breeding,
Applied Microbiology and Biotechnology, Vol.84, No.1, 1-10, Aug. 2009.

(要約): Studies on the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have proceeded in various fields regarding health and dietary requirements in a search for novel and rich sources. Filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, ones reaching 20 g/L and containing 30-70% arachidonic acid as to the total fatty acids. Mutants derived from M. alpina 1S-4, defective in Delta5 and Delta6 desaturases, accumulate triacylglycerols rich in unique PUFAs, i.e., dihomo-gamma-linolenic acid and Mead acid, respectively. Furthermore, various mutants derived from M. alpina 1S-4 have led to the production of oils containing n-1, n-3, n-4, n-6, n-7, and n-9 PUFAs. A variety of genes encoding fatty acid desaturases and elongases involved in PUFA biosynthesis in M. alpina 1S-4 has been isolated and characterized. Molecular breeding of M. alpina strains by means of manipulation of these genes facilitates improvement of PUFA productivity and elucidation of the functions of enzymes involved in PUFA biosynthesis.
(キーワード): Fatty Acid Desaturases / Fatty Acids, Unsaturated / Fungal Proteins / Genetic Engineering / Industrial Microbiology / Mortierella
(出版サイトへのリンク): ● Publication site (DOI): 10.1007/s00253-009-2076-7
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19565237; ● Summary page in Scopus @ Elsevier: 2-s2.0-67649295229

(DOI: 10.1007/s00253-009-2076-7, PubMed: 19565237, Elsevier: Scopus) 櫻谷英治, 安藤晃規, 小川順, 清水昌 :
機能性脂質の微生物による生産. アラキドン酸に関連する油脂の発酵生産を中心として,
蛋白質核酸酵素, Vol.54, No.6, 725-734, 2009年5月.

(キーワード): 油糧微生物 / Mortierella alpina / 発酵油脂 / 高度不飽和脂肪酸 / 代謝工学 / 分子育種
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 19462758; ● CiNii @ 国立情報学研究所 (CRID): 1523669555522006144; ● Search Scopus @ Elsevier (PMID): 19462758

(PubMed: 19462758, CiNii: 1523669555522006144) 小川順, 櫻谷英治, 清水昌 :
機能性脂質の微生物生産,
生物工学会誌, Vol.83, No.7, 339-341, 2005年7月.

(文献検索サイトへのリンク): ● CiNii @ 国立情報学研究所 (CRID): 1520853833398353152

(CiNii: 1520853833398353152) 清水昌, 櫻谷英治, 小川順 :
機能性脂質の微生物生産-アラキドン酸関連高度不飽和脂肪酸含有油脂および共役脂肪酸の生産を中心として-,
オレオサイエンス, Vol.3, No.3, 129-139, 2003年3月.

(出版サイトへのリンク): ● Publication site (DOI): 10.5650/oleoscience.3.129
(文献検索サイトへのリンク): ● Search Scopus @ Elsevier (DOI): 10.5650/oleoscience.3.129

(DOI: 10.5650/oleoscience.3.129) 小川順, 櫻谷英治, 清水昌 :
微生物による機能性脂質の生産,
科学と工業, Vol.76, No.4, 163-170, 2002年4月. 清水昌, 小川順, 櫻谷英治 :
有用油脂生産微生物の代謝工学,
バイオサイエンスとインダストリー, Vol.59, No.7, 451-454, 2001年7月. Milan Certik, Eiji Sakuradani and Sakayu Shimizu :
Desaturase defective fungal mutants: useful tools for the regulation and overproduction of polyunsaturated fatty acids,
Trends in Biotechnol., Vol.16, No.12, 500-505, Dec. 1998.

講演・発表

Eiji Sakuradani, Yoshida Kai, Murakawa Naomi and Takaiku Sakamoto :
Studies on filamentous fungus Fusarium sp. accumulating hydroxy fatty acids,
2022 AOCS Annual Meeting & Expo, May 2022. Eiji Sakuradani, Naomi Murakawa, Konomi Ueno, Takaiku Sakamoto, Yuki Soma, Yoshihiro Izumi, Takeshi Bamba, Akinori Ando, Shigenobu Kishino and Jun Ogawa :
Molecular breeding using a fatty acid hydratase from a filamentous fungus,
The 15th International Symposium on Biocatalysis and Agricultural Biotechnology, Hiroshima, Sep. 2019. Naomi Murakawa, Takaiku Sakamoto and Eiji Sakuradani :
Oxidized fatty acids produced by filamentous fungus Fusarium sp.,
The 1st International Symposium on Chemical Communication(ISCC2019), Tokyo, Jan. 2019. Hiroshi Kikukawa, Eiji Sakuradani, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Metabolic Engineering for Rare PUFA Production by an Oil-producing Fungus Mortierella alpina.,
2018 AOCS Annual Meeting & Expo, Minnesota USA, May 2018. Akinori Ando, Yuki Takemoto, Ryohei Nakatsuji, Shigeru Hiramoto, Eiji Sakuradani and Jun Ogawa :
Practical Eicosapentaenoic Acid (EPA) Production by Mortierella alpina Molecular Breeding nuder Ordinary Temperature.,
2018 AOCS Annual Meeting & Expo, Minnesota USA, May 2018. Eiji Sakuradani, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Production of Various PUFAs by filamentous fungus Mortierella alpina.,
2018 AOCS Annual Meeting & Expo, Minnesota USA, May 2018. Eiji Sakuradani, Naomi Murakawa and Takaiku Sakamoto :
Production of Microbial Lipids using Crude Glycerol.,
2018 AOCS Annual Meeting & Expo, Minnesota USA, May 2018. Akinori Ando, Tomoyo Okuda, Hiroshi Kikukawa, Eiji Sakuradani and Jun Ogawa :
EPA Production by an Oleaginous Fungus Mortierella alpina Breeding at Moderate Temperature,
107th AOCS Annual Meeting & Expo, Utah USA, May 2016. Hiroshi Kikukawa, Eiji Sakuradani, Shigenobu Kishino, Si-Bum Park, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
Characterization of a trifunctional ω3-desaturase from oleaginous fungus Mortierella alpina 1S-4 using a yeast expression system.,
18th Japanese-German Workshop Enzyme Technology, Kyoto, Sep. 2015. Hiroshi Kikukawa, Eiji Sakuradani, Akinori Ando, Sakayu Shimizu and Jun Ogawa :
An Efficient Gene Targeting and Molecular Breeding in Oilproducing Fungus Mortierella alpina with Deletion of lig4 Gene for Non-homologous End Joining,
106th AOCS Annual Meeting & Expo, Utah USA, May 2015. Akinori Ando, Tomoyo Okuda, Hiroshi Kikukawa, Eiji Sakuradani, Jun Shima, Jun Ogawa and Sakayu Shimizu :
Various Rare Polyunsaturated Fatty Acid Productions by Mortierella alpina Breeding.,
106th AOCS Annual Meeting & Expo, Utah, May 2015. J. Ogawa, Eiji Sakuradani, S. Kishino, A. Ando and S. Shimizu :
Fermentative production of polyunsaturated fatty acids and their unique transformation by gut microorganisms,
1st Asian Conference on Oleo Science, Sapporo, Sep. 2014. Hiroshi Kikukawa, Eiji Sakuradani, Shigenobu Kishino, Si-Bum Park, Akinori Ando, Jun Shima, Misa Ochiai, Sakayu Shimizu and Jun Ogawa :
Characterization of a Trifunctional Fatty Acid Desaturase from Oleaginous Filamentous Fungus Mortierella alpina 1S-4 Using a Yeast Expression System,
105th American Oil Chemists' Society Annual Meeting & Expo., Vol.116, No.6, 672-676, San Antonio, May 2014.

(出版サイトへのリンク): ● Publication site (DOI): 10.1016/j.jbiosc.2013.05.023
(文献検索サイトへのリンク): ● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 23790547; ● Summary page in Scopus @ Elsevier: 2-s2.0-84886421743

(DOI: 10.1016/j.jbiosc.2013.05.023, PubMed: 23790547, Elsevier: Scopus) A. Ando, T. Okuda, Eiji Sakuradani, J. Shima, J. Ogawa and S. Shimizu :
Studies of Oleaginous Filamentous Fungus Mortierella alpina for Useful Polyunsaturated Fatty Acid Production,
105th American Oil Chemists' Society Annual Meeting & Expo., San Antonio, May 2014. Eiji Sakuradani, Akinori Ando, Takaiku Sakamoto, Jun Shima, Sakayu Shimizu and Jun Ogawa :
Microbial Production of Various Polyunsaturated Fatty Acids by Molecular Breeding of oleaginous fungus Mortierella alpina,
The 1st International Symposium on Microbial Technology for Food and Energy Security, Bangkok Thailand, Nov. 2013. Eiji Sakuradani, Makiko Naka, Hiroyuki Kanamaru, Yuji Ioka, Masutoshi Nojiri, Jun Ogawa and Sakayu Shimizu :
Operation of n-4 and n-7 pathways for the polyunsaturated fatty acid biosynthesis in a mutant of an arachidonic acid-producing fungus, Mortierella alpina 1S-4,
95th American Oil Chemists' Society Annual Meeting & Expo, in Cincinnati, Ohio, May 2004. 柴田優芽, 茨木暢, 池田汐里, 阪本鷹行, 櫻谷英治 :
フザリウム属糸状菌におけるΔ6 脂肪酸不飽和化酵素遺伝子発現によるγ-リノレン酸生産,
酵素工学研究会第92回講演会, 2024年11月. 池田汐里, 茨木暢, 柴田優芽, 玉野孝一, 阪本鷹行, 櫻谷英治 :
Fusarium属糸状菌における麹菌由来リパーゼ遺伝子過剰発現株の育種,
第23回糸状菌分子生物学コンファレンス, 2024年11月. 柴田優芽, 茨木暢, 池田汐里, 阪本鷹行, 櫻谷英治 :
Fusarium属糸状菌とその近縁種の水酸化脂肪酸生産性評価および形質転換に関する研究,
第23回糸状菌分子生物学コンファレンス, 2024年11月. 茨木暢大, 柴田優芽, 池田汐里, 阪本鷹行, 櫻谷英治 :
Mortierella alpina由来Δ6デサチュラーゼを用いたFusarium sp. 組換え株によるγ-リノレン酸生産,
第23回糸状菌分子生物学コンファレンス, 2024年11月. 櫻谷英治, 柴田優芽, 茨木暢大, 池田汐里, 阪本鷹行 :
廃グリセロールや植物油を利用した Fusarium 属糸状菌による水酸化脂肪酸生産,
第76回日本生物工学会大会, 2024年9月. 久米いずみ, 奥野寧々, 池北愛花, 阪本鷹行, 櫻谷英治 :
Aurantiochytrium属微細藻類を用いたカロテノイド生産条件の検討,
第9回ラビリンチュラシンポジウム, 2024年8月. 大畑陽花, 中川香澄, 竹内道樹, 阪本鷹行, 櫻谷英治 :
藍染液中のインジゴ還元におけるリグニンの役割,
日本農芸化学会中四国支部第67回講演会(例会), 2024年1月. 葭田快, 阪本鷹行, 杉森大助, 櫻谷英治 :
フザリウム属糸状菌による様々な植物油からの水酸化脂肪酸への変換条件の検討,
日本農芸化学会中四国支部第67回講演会(例会), 2024年1月. 葭田快, 阪本鷹行, 杉森大助, 櫻谷英治 :
フザリウム属糸状菌による植物油からの水酸化脂肪酸への変換条件の検討,
酵素工学研究会第90回講演会, 2023年11月. 中川香澄, 松永桃花, 大畑陽花, 阪本鷹行, 櫻谷英治 :
微生物によるインジゴ還元を促進する物質の探索,
2023年度生物工学若手研究者の集い(若手会)夏のセミナー2023, 2023年6月. 迫野眞大, 安藤晃規, 奥田知生, モブライアン, 中辻諒平, 竹本有貴, 池本裕之, 菊川寛史, 阪本鷹行, 櫻谷英治, 小川順 :
Mortierella alpinaによる常温EPA生産を目的とした新規ω3不飽和化酵素の探索および機能解析,
日本農芸化学会2023年度大会, 2023年3月. 佐野壮平, 髙崎廉, 岡久修己, 阪本鷹行, 櫻谷英治 :
香気成分生産性に優れた野生酵母の探索,
日本農芸化学会2022年度中四国支部大会(第63回講演会), 2022年9月. 竹内道樹, 中川香澄, 菊地真由, 宋和慶盛, 安藤晃規, 小川順, 加納健司, 櫻谷英治 :
発酵建て藍染液を科学し，藍文化を育むー電気分析化学手法を基盤としてー,
第82回分析化学討論会(茨城), 2022年5月. 竹内道樹, 中川香澄, 菊地真由, 宋和慶盛, 松永桃花, 阪本鷹行, 安藤晃規, 小川順, 加納健司, 櫻谷英治 :
藍染め染色液の電気化学的解析と微生物燃料電池への応用,
電気化学会第89回大会, 2022年3月. 増田尚輝, 阪本鷹行, 西海信, 渡辺崇人, 櫻谷英治, 三戸太郎, 向井理恵 :
食用昆虫の摂取が実験動物の代謝に及ぼす影響,
日本農芸化学会2022年度大会, 2022年3月. 髙崎廉, 佐野壮平, 岡久修己, 阪本鷹行, 櫻谷英治 :
Non-Saccharomyces酵母変異株の探索とそれらを利用した簡易ビール醸造試験,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 北口直樹, 楠真緒, 木内かなえ, 阪本鷹行, 櫻谷英治 :
担子菌における子実体形成関連遺伝子の探索,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 増田成穂, 池田雄祐, 久米いずみ, 阪本鷹行, 櫻谷英治 :
油糧微生物ラビリンチュラ類の変異によるアスタキサンチン高生産株の育種,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 埴淵滉貴, 前田竜瑚, 阪本鷹行, 櫻谷英治 :
微生物によるデオキシコール酸代謝機構の解析,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 中村悠嗣, 江口主馬, 阪本鷹行, 櫻谷英治 :
油糧微細藻Thraustochytridの変異育種による脂肪酸組成改変,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 吉﨑万莉, 井上朋美, 高野りの, 阪本鷹行, 櫻谷英治 :
Bacillus属細菌による抗菌活性薬剤Iturin生産に関する研究,
日本農芸化学会中四国支部第61回講演会(例会), 2022年1月. 植野美彦, 櫻谷英治, 関陽介, 上岡麻衣子, 浅田元子, 赤松徹也, 宮脇克行, 宇都義浩, 田中保 :
一般選抜後期日程における入学辞退率改善の取り組みーー徳島大学B学部の事例からーー,
第17回大学教育カンファレンスin徳島, 2022年1月. 糸川未来, 安藤晃規, 奥田知生, MO Brian King Himm, 中辻諒平, 竹本有貴, 池本裕之, 菊川寛史, 阪本鷹行, 櫻谷英治, 小川順 :
Mortierella alpina による EPA 生産のための ω3 不飽和化酵素の探索及び機能評価,
2021年度第3回脂質駆動学術産業創生研究部会講演会, 2021年12月. 葭田快, 村川直美, 阪本鷹行, 櫻谷英治 :
水酸化脂肪酸生産性糸状菌Fusarium solani D2株の形質転換条件の検討,
2021年度第3回脂質駆動学術産業創生研究部会講演会, 2021年12月. 増田尚輝, 阪本鷹行, 西海信, 渡辺崇人, 櫻谷英治, 三戸太郎, 向井理恵 :
食用昆虫の脂質組成分析と実験動物への影響,
2021年度第3回脂質駆動学術産業創生研究部会講演会, 2021年12月. 中川香澄, 竹内道樹, 菊池真由, 阪本鷹行, 小川順, 加納健司, 櫻谷英治 :
藍染液中で起こるインジゴの酸化還元に関わる電子伝達メカニズムの解明,
第22回生体触媒化学シンポジウム, 2020年12月. 中川香澄, 竹内道樹, 菊池真由, 阪本鷹行, 小川順, 加納健司, 櫻谷英治 :
藍染液中のインジゴ還元メカニズムの解明,
日本生物工学会西日本支部大会2020(第5回講演会), 2020年11月. 古賀武尊, 阪本鷹行, 櫻谷英治, 田井章博 :
ヒマワリ種子由来のPC12細胞における神経突起形成促進作用物質,
日本農芸化学会中四国支部第57回講演会(例会), 2020年9月. 東洸希, 中村和弘, 阪本鷹行, 櫻谷英治 :
油糧糸状菌のSeipin様タンパク質遺伝子の機能解析,
日本農芸化学会中四国支部第57回講演会(例会), 2020年9月. 北口直樹, 楠真緒, 阪本鷹行, 櫻谷英治 :
アグロバクテリウム法を用いたシイタケ形質転換法の開発,
日本農芸化学会中四国支部第57回講演会(例会), 2020年9月. 島田良美, 下平武彦, 相馬悠希, 和泉自泰, 安藤晃規, 岸野重信, 阪本鷹行, 馬場健史, 小川順, 櫻谷英治 :
油糧糸状菌Mortierella alpinaの分子育種による遊離脂肪酸生産株のリピドーム解析,
日本農芸化学会2020年度大会, 2020年3月. 中川香澄, 多田真奈巳, 阪本鷹行, 竹内道樹, 加納健司, 小川順, 櫻谷英治 :
藍染液の電気化学的解析によるインジゴの測定,
日本農芸化学会2020年度大会, 2020年3月. 村川直美, 阪本鷹行, 上野このみ, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
糸状菌Fusarium solani D2株由来オレイン酸水和酵素の機能解析,
日本農芸化学会中四国支部第56回講演会(例会), 2020年1月. 池田雄祐, 増田成穂, 金崎仁志, 阪本鷹行, 櫻谷英治 :
アスタキサンチン生産性ラビリンチュラの探索および育種,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 中村悠嗣, 江口主馬, 阪本鷹行, 櫻谷英治 :
油糧微生物ラビリンチュラの育種による DHA高生産株の創製,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 坂口悠香, 中川香澄, 髙野仁美, 服部結衣, 阪本鷹行, 櫻谷英治 :
海洋生物共生菌によるアルキルグリセロール生産,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 吉﨑万莉, 井上朋美, 阪本鷹行, 櫻谷英治 :
灰色かび病菌Botrytis cinerea に抗菌特性を示す微生物の探索,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 西川和毅, 埴淵滉貴, 阪本鷹行, 櫻谷英治 :
コール酸資化性菌におけるコール酸代謝経路の解析,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 上野このみ, 村川直美, 阪本鷹行, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
油糧糸状菌Mortierella alpinaの育種による10-ヒドロキシステアリン酸生産,
日本農芸化学会 2019年度西日本・中四国支部合同大会, 2019年11月. 中村和弘, 東洸希, 阪本鷹行, 島田良美, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
油糧糸状菌Mortierella alpinaにおけるPerilipin様タンパク質の機能解析,
第19回糸状菌分子生物学コンファレンス, 2019年11月. 阪本鷹行, 村川直美, 上野このみ, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
糸状菌Fusarium solani D2株から単離した脂肪酸水和酵素の機能解析,
第19回糸状菌分子生物学コンファレンス, 2019年11月. 野口愛佳, 村川直美, 上野このみ, 阪本鷹行, 櫻谷英治 :
水酸化脂肪酸生産性糸状菌Fusarium solani D2株の形質転換法の開発,
第19回糸状菌分子生物学コンファレンス, 2019年11月. 松家彩恵, 阪本鷹行, 岡久修己, 櫻谷英治 :
自然界より単離した酵母の醸造特性評価,
日本農芸化学会2019年度大会, 2019年3月. 村川直美, 上野このみ, 阪本鷹行, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
糸状菌由来オレイン酸水和酵素遺伝子の機能解析,
日本農芸化学会2019年度大会, 2019年3月. 中川香澄, 多田真奈巳, 西林菜華, 阪本鷹行, 清藤鈴奈, 竹内道樹, 加納健司, 小川順, 櫻谷英治 :
藍染液中の電子伝達に関わる微生物・反応促進物質の探索,
日本農芸化学会2019年度大会, 2019年3月. 島田良美, 岸野重信, 油屋駿介, 青木航, 植田充美, 安藤晃規, 馬場健史, 小川順, 櫻谷英治 :
油糧糸状菌Mortierella alpinaの漏出脂質プロテオーム解析と漏出株の創製,
日本農芸化学会2019年度大会, 2019年3月. 櫻谷英治 :
真核微生物がつくる多様な機能性脂質,
第13回日本ゲノム微生物学会年会ー日本農芸化学会共済シンポジウム, 2019年3月. 中村悠嗣, 阪本鷹行, 櫻谷英治 :
油糧微生物ラビリンチュラにおけるDHA生産条件の検討,
日本農芸化学会中四国支部第52回講演会, 2018年9月. 大塚拓実, 阪本鷹行, 櫻谷英治 :
植物寄生菌の脂質解析,
日本農芸化学会中四国支部第52回講演会, 2018年9月. 多田真奈巳, 中川香澄, 阪本鷹行, 清藤鈴奈, 竹内道樹, 小川順, 櫻谷英治 :
藍染液より単離した微生物の解析,
日本農芸化学会中四国支部第52回講演会, 2018年9月. 櫻谷英治, 小川慶徳 :
徳島大学ビール(Very Ale.)の開発,
日本醸造学会若手の会第32回スチューデントサイエンティストプログラム, 2018年9月. 櫻谷英治 :
さまざまな生物資源から単離された微生物の活用,
第70回日本生物工学会大会, 2018年9月. 清藤鈴奈, 竹内道樹, 久郷将見, 多田真奈巳, 阪本鷹行, 櫻谷英治, 小川順 :
インジゴ還元菌におけるメディエーターの影響とその機能解析,
日本農芸化学会2018年度大会, 2018年3月. 村川直美, 加納みずほ, 阪本鷹行, 安藤晃規, 岸野重信, 相馬悠希, 馬場健史, 小川順, 櫻谷英治 :
廃グリセロール資化性糸状菌が生産する脂肪酸代謝産物,
日本農芸化学会2018年度大会, 2018年3月. 島田良美, 安藤晃規, 岸野重信, 櫻谷英治, 小川順 :
油糧糸状菌Mortierella alpinaの分子育種による遊離脂肪酸生産に関する検討,
日本農芸化学会2018年度大会, 2018年3月. 櫻谷英治 :
藍染工程のインジゴ還元に関わる微生物群について,
Visionary 農芸化学100 シンポジウム, 2017年12月. 松家彩恵, 阪本鷹行, 櫻谷英治 :
エタノール生産性天然酵母の単離および諸性質解明,
第9回日本醸造学会若手シンポジウム, 2017年10月. 奥田知生, 安藤晃規, 櫻谷英治, 鎌田望, 落合美佐, 小川順 :
糸状菌 Mortierella alpina 1S-4 株における誘導発現プロモーターの探索と評価,
日本農芸化学会関西・中四国・西日本支部 2017年度合同大阪大会(第49回講演会), 2017年9月. 阪本鷹行, 太田里咲, 楠美波, 高野仁美, 櫻谷英治 :
スクアレン生産性酵母の探索と培養条件の検討,
日本農芸化学会関西・中四国・西日本支部 2017年度合同大阪大会(第49回講演会), 2017年9月. 加納みずほ, 亀川優一, 栗田千波, 阪本鷹行, 櫻谷英治 :
廃グリセロール資化性菌による微生物油脂生産,
日本農芸化学会関西・中四国・西日本支部 2017年度合同大阪大会(第49回講演会), 2017年9月. 上田堅人, 大塚拓実, 阪本鷹行, 櫻谷英治 :
冬虫夏草の脂質解析,
日本農芸化学会関西・中四国・西日本支部 2017年度合同大阪大会(第49回講演会), 2017年9月. 松家彩恵, 阪本鷹行, 櫻谷英治 :
自然界より単離したエタノール生産性酵母の諸性質解明,
日本農芸化学会中四国支部第48回講演会, 2017年6月. 清藤鈴奈, 竹内道樹, 久郷将見, 四方田和弥, 阪本鷹行, 櫻谷英治, 小川順 :
インジゴ還元能の定量的評価法を用いたインジゴ還元菌の探索及び機能解析,
日本農芸化学会中四国支部第48回講演会, 2017年6月. 久郷将見, 竹内道樹, 四方田和弥, 阪本鷹行, 櫻谷英治, 小川順 :
インジゴ還元菌の探索及び定量的評価法を用いる機能解析,
日本農芸化学会2017年度大会, 2017年3月. 井出紗奈江, 阪本鷹行, 安藤晃規, 岸野重信, 和泉自泰, 馬場健史, 島純, 小川順, 櫻谷英治 :
油糧糸状菌 Mortierella alpina から誘導したリシノール酸生産変異株について,
日本農芸化学会2017年度大会, 2017年3月. 高野仁美, 阪本鷹行, 櫻谷英治 :
海洋性微生物のジピコリン酸生産性評価,
日本農芸化学会2017年度大会, 2017年3月. 奥田知生, 安藤晃規, 櫻谷英治, 鎌田望, 落合美佐, 小川順 :
油糧微生物 Mortierella alpina 1S-4株の分子育種に有用なプロモーターの探索および評価,
日本農芸化学会2017年度大会, 2017年3月. 島田良美, 安藤晃規, 和泉自泰, 馬場健史, 岸野重信, 櫻谷英治, 島純, 小川順 :
TCA回路酵素遺伝子過剰発現が Mortierella alpina の脂質生産性に及ぼす影響,
日本農芸化学会2017年度大会, 2017年3月. 奥田知生, 安藤晃規, 櫻谷英治, 鎌田望, 落合美佐, 小川順 :
油糧微生物Mortierella alpina 1S-4株における遺伝子発現情報に基づいたプロモーターの探索および解析,
第7回学際的脂質創生研究部会講演会, 2017年1月. 櫻谷英治, 阪本鷹行, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 島純, 小川順 :
リシノール酸生産を目指した油糧微生物 Mortierella alpina の分子育種,
第7回学際的脂質創生研究部会講演会, 2017年1月. 和泉自泰, 安藤晃規, 阪本鷹行, 岸野重信, 相馬悠希, 福崎英一郎, 櫻谷英治, 島純, 小川順, 馬場健史 :
メタボローム解析による油糧微生物の脂質生産性向上のための代謝改変戦略の立案,
第7回学際的脂質創生研究部会講演会, 2017年1月. 岡直宏, 阪本鷹行, 櫻谷英治, 團昭紀, 濵野龍夫 :
有用海藻Agardhiella subulataの脂肪酸分析について,
第3回日本生物工学会西日本支部講演会, 2016年12月. 阪本鷹行, 森下賢志, 富永康子, 濱野百花, 安藤晃規, 岸野重信, 和泉自泰, 馬場健史, 島純, 小川順, 櫻谷英治 :
油糧糸状菌Mortierella alpinaの分子育種によるリシノール酸生産,
第3回日本生物工学会西日本支部講演会, 2016年12月. 梶浦雅斗, 加納みずほ, 亀川優一, 栗田千波, 阪本鷹行, 櫻谷英治 :
廃グリセロール資化性菌の探索と微生物油脂生産への応用,
第3回日本生物工学会西日本支部講演会, 2016年12月. 四方田和弥, 阪本鷹行, 尾下竜次, 久郷将見, 竹内道樹, 小川順, 櫻谷英治 :
インジゴ還元に関わる微生物の探索及び機能性評価,
第3回日本生物工学会西日本支部講演会, 2016年12月. 富永康子, 濱野百花, 阪本鷹行, 井出紗奈江, 菊川寛史, 安藤晃規, 岸野重信, 和泉自泰, 馬場健史, 島純, 小川順, 櫻谷英治 :
リシノール酸高生産株の作製とその脂質解析,
第16回糸状菌分子生物学コンファレンス, 2016年11月. 阪本鷹行, 富永康子, 井出紗奈江, 奥田知生, 安藤晃規, 岸野重信, 和泉自泰, 馬場健史, 島純, 櫻谷英治, 小川順 :
油糧糸状菌におけるリシノール酸生産株の分子育種,
第16回糸状菌分子生物学コンファレンス, 2016年11月. 亀川優一, 加納みずほ, 栗田千波, 阪本鷹行, 櫻谷英治 :
廃グリセロールを利用した微生物油脂生産,
日本農芸化学会2016年度中四国支部大会, 2016年9月. 菊川寛史, 櫻谷英治, 安藤晃規, 小川順 :
油糧微生物Mortierella alpinaの標的遺伝子破壊による希少脂肪酸生産,
日本農芸化学会2016年度大会, 2016年3月. 島田良美, 安藤晃規, 和泉自泰, 馬場健史, 岸野重信, 櫻谷英治, 島純, 小川順 :
TCA回路酵素遺伝子過剰発現によるMortierella alpinaの脂質生産性向上の検討,
日本農芸化学会2016年度大会, 2016年3月. 菊川寛史, 櫻谷英治, 安藤晃規, 小川順 :
油脂生産性糸状菌Mortierella alpinaの代謝改変による希少脂肪酸生産,
第6回学際的脂質創生研究部会講演会, 2016年1月. 安藤晃規, 奥田知生, 櫻谷英治, 小川順 :
油脂生産性糸状菌Mortierella alpinaによるオメガ3脂肪酸の生産,
第6回学際的脂質創生研究部会講演会, 2016年1月. 亀川優一, 栗田千波, 阪本鷹行, 櫻谷英治 :
グリセロールを利用した微生物油脂生産,
第6回学際的脂質創生研究部会講演会, 2016年1月. 菊川寛史, 村椿達哉, 安藤晃規, 櫻谷英治, 清水昌, 小川順 :
糸状菌Mortierella chlamydosporaの脂肪酸鎖長延長酵素遺伝子の機能解析,
第67回日本生物工学会大会, 2015年10月. 浅井大貴, 櫻谷英治, 菊川寛史, 安藤晃規, 小川順 :
EPA の常温発酵生産を目的とした油糧微生物Mortierella alpina 1S-4におけるPythium sulcatum由来ω3不飽和化酵素遺伝子の過剰発現,
第67回日本生物工学会大会, 2015年10月. 竹本有貴, 中辻諒平, 櫻谷英治, 安藤晃規, 平本茂, 原田昌卓, 小川順 :
酵母発現系を活用した糸状菌 Pythium sulcatum NBRC100117 株由来 ω3 不飽和化酵素の機能解析,
第67回日本生物工学会大会, 2015年10月. 和泉自泰, 安藤晃規, 阪本鷹行, 光永均, 中尾素直, 岸野重信, 福崎英一郎, 櫻谷英治, 島純, 小川順, 馬場健史 :
油糧糸状菌Mortierella alpinaのメタボローム解析,
第67回日本生物工学会大会, 2015年10月. 和泉自泰, 安藤晃規, 阪本鷹行, 光永均, 中尾素直, 岸野重信, 福崎英一郎, 櫻谷英治, 島純, 小川順, 馬場健史 :
油糧微生物 Mortierella alpinaのメタボローム解析,
第9回メタボロームシンポジウム, 2015年10月. 浅井大貴, 中辻諒平, 安藤晃規, 櫻谷英治, 小川順 :
糸状菌 Pythium sulcatum 由来ω3 不飽和化酵素遺伝子の機能解析および油糧微生物 Mortierella alpina 1S-4 における過剰発現,
2015年度日本農芸化学会中部・関西支部合同大会, 2015年9月. 湯浅雄介, 阪本鷹行, 清水昌, 小川順, 櫻谷英治 :
ステロール生産性 Mortierella alpina の形質転換系の確立,
日本農芸化学会中四国支部第42回講演会, 2015年6月. 菊川寛史, 中辻諒平, 浅井大貴, 櫻谷英治, 安藤晃規, 小川順 :
油糧微生物Mortierella alpinaへのPythium sp.由来ω3不飽和化酵素遺伝子の導入によるエイコサペンタエン酸の常温生産,
日本農芸化学会2015年度大会, 2015年3月. 盤若明日香, 菊川寛史, 安藤晃規, 櫻谷英治, 落合美佐, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4株におけるΔ12不飽和化酵素遺伝子破壊によるミード酸高生産株の構築,
日本農芸化学会2015年度大会, 2015年3月. 阪本鷹行, 菊川寛史, 安藤晃規, 櫻谷英治, 落合美佐, 島純, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4株におけるΔ6不飽和化酵素遺伝子をターゲットとした分子育種株の構築,
日本農芸化学会2015年度大会, 2015年3月. 奥田知世, 安藤晃規, 櫻谷英治, 鎌田望, 落合美佐, 小川順 :
油糧性微生物Mortierella alpina 1S-4株における新規プロモーターの探索および評価,
日本農芸化学会2015年度大会, 2015年3月. 菊川寛史, 中辻諒平, 浅井大貴, 櫻谷英治, 安藤晃規, 小川順 :
糸状菌 Pythium sp. 由来 ω3 不飽和化酵素遺伝子の酵母発現系を用いた機能解析,
第14回糸状菌分子生物学コンファレンス, 2014年11月. 安藤晃規, 中辻諒平, 菊川寛史, 櫻谷英治, 小川順 :
Pythium sp.由来 ω3 不飽和化酵素遺伝子を活用した油糧微生物 Mortierella alpina 1S-4 による EPA の常温生産,
第14回糸状菌分子生物学コンファレンス, 2014年11月. 阪本鷹行, 櫻谷英治, 安藤晃規, 島純, 小川順 :
オレイン酸及びリノール酸高生産を目的としたMortierella alpinaの分子育種,
第14回糸状菌分子生物学コンファレンス, 2014年11月. 菊川寛史, 櫻谷英治, 安藤晃規, 落合美佐, 清水昌, 小川順 :
油脂生産性糸状菌Mortierella alpina 1S-4の脂肪酸鎖長延長酵素遺伝子破壊による脂肪酸組成の改変,
2014年度日本農芸化学会関西支部大会(第486回講演会), 2014年9月. 浅井大貴, 櫻谷英治, 中辻諒平, 安藤晃規, 菊川寛史, 小川順 :
糸状菌Pythium sp. 由来ω3不飽和化酵素の酵母発現系を用いた機能解析,
第66回日本生物工学会大会, 2014年9月. 中辻諒平, 櫻谷英治, 安藤晃規, 菊川寛史, 小川順 :
油糧微生物Mortierella alpina 1S-4を活用したPythium sp. 由来ω3 不飽和化酵素遺伝子導入による常温EPA生産,
第66回日本生物工学会大会, 2014年9月. 菊川寛史, 櫻谷英治, 安藤晃規, 落合美佐, 清水昌, 小川順 :
油脂生産性微生物Mortierella alpina 1S-4 の脂肪酸鎖長延長酵素遺伝子破壊による脂肪酸組成の改変,
第66回日本生物工学会大会, 2014年9月. 奥田知生, 安藤晃規, 櫻谷英治, 鎌田望, 落合美佐, 小川順 :
油糧性糸状菌Mortierella alpina 1S-4株の分子育種に有用な新規プロモーターの探索と評価,
第66回日本生物工学会大会, 2014年9月. 安藤晃規, 奥田知生, 根来宏明, 菊川寛史, 櫻谷英治, 島純, 清水昌, 小川順 :
油糧微生物Mortierella alpinaの分子育種によるエイコサテトラエン酸(ETA)の常温生産,
第66回日本生物工学会大会, 2014年9月. 阪本鷹行, 櫻谷英治, 安藤晃規, 島純, 小川順 :
Mortierella alpinaにおけるオレイン酸及びリノール酸高生産株の分子育種,
第66回日本生物工学会大会, 2014年9月. 阪本鷹行, 櫻谷英治, 安藤晃規, 島純, 小川順 :
オレイン酸及びリノール酸高蓄積性Mortierella alpinaの育種,
生物工学若手研究者の集い, 2014年7月. 菊川寛史, 櫻谷英治, 安藤晃規, 清水昌, 小川順 :
油脂生産性糸状菌Moltierella alpinaの標的遺伝子破壊系の構築と希少脂質生産,
生物工学若手研究者の集い, 2014年7月. 櫻谷英治, 安藤晃規, 岸野重信, 日比慎, 小川順 :
脂質バランス栄養食品を創出する新規オメガ3脂肪酸素材の開発,
日本農芸化学会 2014年度大会, 2014年3月. 阪本鷹行, 櫻谷英治, 安藤晃規, 島純, 小川順 :
油糧微生物Mortierella alpinaにおけるオレイン酸及びリノール酸生産性の向上,
日本農芸化学会2014年度大会, 2014年3月. 阪本鷹行, 櫻谷英治, 安藤晃規, 島純, 小川順 :
Mortierella alpinaの分子育種によるオレイン酸生産,
第4回学際的脂質創生研究部会講演会, 2014年1月. 菊川寛史, 浅岡卓也, 櫻谷英治, 奥田知生, 安藤晃規, 阪本鷹行, 島純, 小川順 :
油糧微生物Mortierella alpina 1S-4における外来不飽和化酵素遺伝子発現による高度不飽和脂肪酸生産,
第65回日本生物工学会大会, 2013年9月. 櫻谷英治, 村椿達哉, 西馬場由美, 菊川寛史, 安藤晃規, 小川順 :
油糧微生物Mortierella chlamydosporaにおける脂肪酸鎖長延長酵素遺伝子の機能解析,
日本農芸化学会2013年大会, 2013年3月. 櫻谷英治, 西馬場由美, 菊川寛史, 安藤晃規, 小川順 :
Mortierella属糸状菌によるエイコセン酸生産,
日本農芸化学会2013年大会, 2013年3月. 櫻谷英治, 三宅貴士, 落合美佐, 安藤晃規, 小川順 :
油糧微生物Mortierella alpina 1S-4のcaleosin遺伝子の単離と過剰発現による脂質蓄積に及ぼす効果,
日本農芸化学会2013年大会, 2013年3月. 浅岡卓也, 奥田知生, 櫻谷英治, 安藤晃規, 小川順 :
微細藻由来Δ6不飽和化酵素遺伝子発現による脂質生産性糸状菌Mortierella alpinaの脂肪酸組成改変,
日本農芸化学会2013年大会, 2013年3月. 落合美佐, 櫻谷英治, 安藤晃規, 小川順 :
油糧微生物Mortierella alpina 1S-4由来phospholipid:diacylglycerol acyltransferaseホモログの機能解析,
日本農芸化学会2013年大会, 2013年3月. 藤井茂貴, 櫻谷英治, 田中晃一, 小川順, 島純 :
酵母のストレス耐性強化に向けたエルゴステロール蓄積増強に寄与する生合成遺伝子の探索,
日本農芸化学会2013年大会, 2013年3月. 網干貴子, 中嶋祐二, 清水伸泰, 本田善之, 櫻谷英治, 小川順, 西田律夫, 桑原保正, 天野洋, 森直樹 :
コナダニ類における 9,12-octadecadienoic acid の生合成,
日本農芸化学会2013年大会, 2013年3月. 浅岡卓也, 櫻谷英治, 安藤晃規, 落合美佐, 小川順 :
アラキドン酸生産性糸状菌Mortierella alpina 1S-4由来アシルCoA合成酵素遺伝子の機能解析,
第3回学際的脂質創生研究部会講演会, 2013年1月. 浅岡卓也, 櫻谷英治, 安藤晃規, 落合美佐, 小川順 :
油糧微生物Mortierella alpina 1S-4 由来アシルCoA 合成酵素遺伝子の機能解析,
第64回日本生物工学会大会, 2012年10月. 安藤晃規, 根来宏明, 奥田知生, 菊川寛史, 櫻谷英治, 島純, 清水昌, 小川順 :
外来Δ17不飽和化酵素遺伝子導入による油糧微生物Mortierella alpinaでの常温EPA生産,
日本農芸化学会2012年大会, 2012年3月. 櫻谷英治, 浅岡卓也, 安藤晃規, 落合美佐, 小川順 :
油糧微生物Mortierella alpina 1S-4由来アシルCoA合成酵素遺伝子の過剰発現による脂質生産性向上,
日本農芸化学会2012年大会, 2012年3月. 中谷政人, 櫻谷英治, 菊川寛史, 安藤晃規, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4のku80遺伝子破壊株の作製,
日本農芸化学会2012年大会, 2012年3月. 奥田知生, 安藤晃規, 櫻谷英治, 小川順 :
海洋微生物Crypthecodinium sp. D31株によるドコサヘキサエン酸を含有するリン脂質の生産,
日本農芸化学会2012年大会, 2012年3月. 村上武, 島田良美, 櫻谷英治, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4に由来するSterol acyltransferaseホモログ遺伝子のクローニングと機能解析,
日本農芸化学会2012年大会, 2012年3月. 島田良美, 櫻谷英治, 安藤晃規, 清水昌, 小川順 :
油糧微生物Mortierella alpinaの分子育種による希少ステロール生産(2),
日本農芸化学会2012年大会, 2012年3月. 島田良美, 櫻谷英治, 清水昌, 小川順 :
油糧微生物Mortierella alpinaの分子育種による希少ステロール生産(1),
日本農芸化学会2012年大会, 2012年3月. 菊川寛史, 櫻谷英治, 安藤晃規, 落合美佐, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4に由来するω3脂肪酸不飽和化酵素の機能性評価,
第2回学際的脂質創生研究部会講演会, 2012年1月. 奥田知生, 西馬場由美, 安藤晃規, 櫻谷英治, 小川順 :
海洋性微生物Crypthecodinium sp. D31 株によるドコサヘキサエン酸生産に影響を及ぼす培養条件の検討,
第63回日本生物工学会大会, 2011年9月. 村上武, 櫻谷英治, 福岡義宏, 島田良美, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4由来ERG28ホモログ遺伝子のクローニングと機能解析,
第63回日本生物工学会大会, 2011年9月. 櫻谷英治, 菊川寛史, 安藤晃規, 落合美佐, 小川順 :
油糧微生物Mortierella alpina 1S-4由来ω3不飽和化酵素の機能解析,
第63回日本生物工学会大会, 2011年9月. 安藤晃規, 辻村郁子, 田中ゆか, 菊川寛史, 奥田知生, 櫻谷英治, 島純, 清水昌, 小川順 :
ω3系脂肪酸の特異的生産を目的とした油糧微生物Mortierella alpinaの分子育種,
第63回日本生物工学会大会, 2011年9月. 安藤晃規, 田中ゆか, 菊川寛史, 奥田知生, 櫻谷英治, 島純, 清水昌, 小川順 :
ω3系脂肪酸生産を目的とした高度不飽和脂肪酸生産性糸状菌Mortierella alpinaの分子育種,
日本農芸化学会2011年大会, 2011年3月. 奥田知生, 西馬場由美, 安藤晃規, 櫻谷英治, 小川順 :
海洋性微生物Crypthecodinium sp. D31株を用いたドコサヘキサエン酸生産条件の検討,
日本農芸化学会2011年大会, 2011年3月. 櫻谷英治, 福岡義宏, 村上武, 島田良美, 清水昌, 小川順 :
油糧微生物Mortierella alpina 1S-4に由来するERG28ホモログ遺伝子のクローニングと機能解析,
日本農芸化学会2011年大会, 2011年3月. 落合美佐, 櫻谷英治, 安藤晃規, 小川順 :
高度不飽和脂肪酸生産菌Mortierella alpina由来membrane bound O-acyltransferase,
日本農芸化学会2011年大会, 2011年3月. 西馬場由美, 櫻谷英治, 奥田知生, 安藤晃規, 小川順 :
油糧海洋性微生物由来Δ4不飽和化酵素及びΔ5鎖長延長酵素ホモログ遺伝子の機能解析,
第1回学際的脂質創生研究部会講演会, 2011年1月. 長尾慧, 櫻谷英治, 小林宏治, 安藤晃規, 小川順 :
アルカン資化性微細藻Prototheca zopfiiのsecondary alcohol dehydrogenaseの精製と遺伝子クローニング,
第1回学際的脂質創生研究部会講演会, 2011年1月. 安藤晃規, 田中ゆか, 奥田知生, 櫻谷英治, 島純, 小川順, 清水昌 :
油糧微生物Mortierella alpinaの多重栄養要求性変異株の構築と諸性質の評価,
第10回糸状菌分子生物学コンファレンス, 2010年11月. 櫻谷英治, 長尾慧, 小林宏治, 小川順 :
Prototheca zopfii JCM9400 由来secondary alcohol dehydrogenase の精製とクローニング,
第62回日本生物工学会大会, 2010年10月. 西馬場由美, 櫻谷英治, 奥田知生, 安藤晃規, 小川順 :
海洋性油糧微生物より単離した脂肪酸不飽和化酵素ホモログ遺伝子の解析,
第62回日本生物工学会大会, 2010年10月. 櫻谷英治, 奥田知生, 安藤晃規, 小川順 :
ドコサヘキサエン酸生産性に着目した海洋性微生物の単離と評価,
第62回日本生物工学会大会, 2010年10月. 安藤晃規, 田中ゆか, 奥田知生, 櫻谷英治, 島純, 小川順 :
高度不飽和生産性糸状菌Mortierella alpina 1S-4 株の二重栄養要求性変異株の取得,
第62回日本生物工学会大会, 2010年10月.

研究会・報告書: 阪本鷹行, 村川直美, 上野このみ, 安藤晃規, 岸野重信, 相馬悠希, 和泉自泰, 馬場健史, 小川順, 櫻谷英治 :
糸状菌に由来する脂肪酸水和酵素の機能解析,
酵素・補酵素研究会2019, 2019年7月. 櫻谷英治, 村川直美, 阪本鷹行 :
糸状菌Fusarium sp.由来オレイン酸水和酵素遺伝子の機能解析,
化学コミュニケーションのフロンティア第5回公開シンポジウム, 2019年6月. 植野美彦, 関陽介, 矢部拓也, 米村重信, 阪上浩, 生島仁史, 藤猪英樹, 白山靖彦, 山田健一, 木下和彦, 櫻谷英治, 古屋 S. 玲, 上岡麻衣子 :
令和2年度徳島大学高等教育研究センターアドミッション部門報告書,
令和2年度徳島大学高等教育研究センターアドミッション部門報告書, 2021年3月.

特許: 河島洋, 櫻谷英治, 小川順, 清水昌 : 1種類の高度不飽和脂肪酸残基3個から成るトリグリセライドの製造方法，およびその利用, 特願2009-269167 (2009年12月), 特開2010-42037 (2010年2月), . 瀧村靖, 清水昌, 櫻谷英治 : 2-フェニルエチルアルコールの製造方法, 特願2007-288703 (2007年10月), 特開2009-112245 (2009年5月), . 瀧村靖, 清水昌, 櫻谷英治 : ケトン体及び/又は2級アルコールの製造法, 特願2006-342355 (2006年12月), 特開2008-148663 (2008年7月), . 清水昌, 櫻谷英治 : 脂肪酸鎖長延長酵素遺伝子及びその利用, 特願2006-291255 (2006年10月), 特開2008-73030 (2008年4月), . 清水昌, 櫻谷英治 : ω3脂肪酸不飽和化活性を有するポリペプチドおよびそのポリペプチドをコードするポリヌクレオチドならびにそれらの利用, 特願2004-241671 (2004年8月), 特開2006-55104 (2006年3月), 特許第4587451号 (2010年9月). 落合美佐, 河島洋, 清水昌, 櫻谷英治 : 脂質生産菌の育種方法およびその利用, 特願2004-107512 (2004年3月), 特開2005-287403 (2005年10月), 特許第4481702号 (2010年3月).
作品: 研究者総覧に該当データはありませんでした。
補助金・競争的資金: フザリウム属糸状菌の酸化型脂質蓄積機構の解明と機能性脂質生産への応用 (研究課題/領域番号: 23K26834 )
藍染の「発酵建て」から導く強アルカリ微生物燃料電池の開発 (研究課題/領域番号: 21K19868 )
植物-微生物間の化学コミュニケーションを担う新たな脂質リガンドの探索 (研究課題/領域番号: 20H04779 )
フザリウム属糸状菌がつくる生理活性脂質リガンドの探索 (研究課題/領域番号: 18H04623 )
海洋生物の腸管に共生する海洋性微生物を利用した希少脂質生産法の開発 (研究課題/領域番号: 17K07723 )
高温高アルカリ環境で機能する新規微生物酸化還元・電子伝達機能の探索と開発 (研究課題/領域番号: 16K14884 )
長鎖アルカンの合成と分解に関わる酵素群の探索と脂質変換反応への応用 (研究課題/領域番号: 24658078 )
新規脂質変換反応の探索と機能性脂質生産プロセスの開発 (研究課題/領域番号: 22380051 )
微生物における還元的脂肪酸・有機酸変換機能の探索と開発 (研究課題/領域番号: 20380050 )
油糧微生物の分子育種と微生物の特異な代謝機能を活用した有用物質生産 (研究課題/領域番号: 19688006 )
非侵襲的細胞脂質解析を目指した生体触媒モジュールの開発 (研究課題/領域番号: 19021027 )
窒素含有キラル化合物不斉合成のための微生物生産プロセスの開発 (研究課題/領域番号: 18380057 )
微生物機能の多面的活用による新規機能性脂質の創製 (研究課題/領域番号: 18208009 )
細胞脂質解析のための生体触媒モジュールの開発 (研究課題/領域番号: 18038026 )
Single Cell Oilの新展開「機能性脂肪酸から構造脂質・機能性脂質へ」 (研究課題/領域番号: 17658040 )
高度不飽和脂肪酸蓄積性糸状菌の遺伝子組み換え技術の確立と生産への応用 (研究課題/領域番号: 16780054 )
新規な酵素法による2つのキラル中心を持つ化合物の立体選択的生産 (研究課題/領域番号: 16380060 )
研究者番号（10362427）による検索

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2024年11月22日更新

専門分野・研究分野: ライフサイエンス (Life sciences) [応用生物化学 (Applied biochemistry)]
所属学会・所属協会: 日本醸造学会
日本生物工学会
社団法人日本農芸化学会
糸状菌分子生物学研究会
一般財団法人バイオインダストリー協会
酵素工学研究会
Applied Microbiology and Biotechnology
委員歴・役員歴: 日本醸造学会 (若手の会運営委員 [2013年10月〜2016年10月])
日本生物工学会 (若手会役員 [2005年4月〜2007年3月], 西日本支部庶務 [2015年4月〜2017年3月], 西日本支部評議員 [2015年4月〜2017年3月])
社団法人日本農芸化学会 (代議員 [2010年4月〜2012年3月], 広報委員 [2015年6月〜2019年5月], 学術活動強化委員会委員 [2015年6月〜2021年5月], 中四国支部副支部長 [2017年6月〜2019年5月], 中四国支部支部長 [2019年6月〜2021年5月], 大会WG 委員 [2019年6月〜2021年5月], 学術活動強化委員会委員長 [2021年6月〜2023年5月])
糸状菌分子生物学研究会 (運営委員 [2015年7月〜2017年6月])
Applied Microbiology and Biotechnology (Editorial Board Members [2012年1月〜2013年12月])
受賞: 2007年6月, 酵素応用シンポジウム研究奨励賞 (天野エンザイム)
2009年3月, 農芸化学奨励賞 (社団法人日本農芸化学会)
2011年9月, 酵素工学研究奨励賞 (酵素工学研究会)
2022年3月, 2022年度電気化学会論文賞 (公益社団法人電気化学会)
2023年3月, 2022年Bioscience, Biotechnology, and Biochemistry(BBB) 論文賞 (社団法人日本農芸化学会)
活動: 広報委員会委員長 (2016年4月〜2017年3月)
教務委員会副委員長 (2017年4月〜2018年3月)
インターンシップ委員会副委員長 (2017年4月〜2018年3月)
教務委員会委員長 (2018年4月〜2019年3月)
インターンシップ委員会委員長 (2018年4月〜2019年3月)
入試委員会副委員長 (2019年5月〜2020年4月)
食料科学コース長 (2020年4月〜2022年3月)

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2024年11月16日更新

研究者番号: 10362427

所属（現在）: 2024/4/1 : 徳島大学, 大学院社会産業理工学研究部(生物資源産業学域), 教授

所属（過去の研究課題 情報に基づく）*注記: 2017/4/1 – 2024/4/1 : 徳島大学, 大学院社会産業理工学研究部(生物資源産業学域), 教授
2016/4/1 : 徳島大学, 大学院生物資源産業学研究部, 教授
2014/4/1 : 徳島大学, ソシオテクノサイエンス研究部, 教授
2012/4/1 – 2013/4/1 : 京都大学, (連合)農学研究科(研究院), 助教
2012/4/1 : 京都大学, 大学院・農学研究科, 助教
2007/4/1 – 2011/4/1 : 京都大学, 農学研究科, 助教
2010/4/1 : 京都大学, 大学院・農学研究科, 助教
2004/4/1 – 2006/4/1 : 京都大学, 農学研究科, 助手

審査区分/研究分野

研究代表者

生物系 / 農学 / 農芸化学 / 応用微生物学
理工系
中区分64:環境保全対策およびその関連分野
小区分38030:応用生物化学関連

研究代表者以外

生物系 / 農学 / 農芸化学 / 応用微生物学
理工系

キーワード

研究代表者

アラキドン酸 / 高度不飽和脂肪酸 / 宿主ベクター系 / モルティエレラ / 糸状菌 / RNAi / モルティエラ / 脂肪酸鎖長延長酵素 / 発酵生産 / 分子育種 / アルカン / デスモステロール / 脂肪酸不飽和化酵素 / プロトテカ / ステロール / 油脂発酵 / Mortierella alpina / 発酵 / 微生物油脂 / エイコサペンタエン酸 / 油糧微生物 / 相同組み換え / 相同組換え / 微細藻 / サブターミナル酸化 / Prototheca / alkane degradation / subterminal oxidation / secondary alcohol / 共生微生物 / 脂質 / 微生物 / バイオテクノロジー / 脂肪酸二次代謝産物 / 水酸化脂肪酸 / フザリウム / hydroxy fatty acid / Fusarium / lipid / ligand / インジゴ / 藍染 / 微生物燃料電池 / サイクリックボルタンメトリー / 酸化還元酵素 / フザリウム属糸状菌 / 顕微ラマン分光

研究代表者以外

Single Cell 0il / 構造脂質 / トリアシルグリセロール / Mortierella alpina / ジアシルグリセロール / 高度不飽和脂肪酸 / リパーゼ / triarachidonyl glycerol / Single cell oil / アラキドン酸 / 脂質 / P450 / ペルオキシダーゼ / 脂肪酸 / 変異型酵素 / 過酸化水素 / NADPH / 発酵生産 / 共役リノール酸 / 乳酸菌 / デスモステロール / 共役高度不飽和脂肪酸 / Clostridium / 嫌気的反応 / 脂肪酸の飽和化 / Mortierella / 分子育種 / 変異株 / 宿主ベクター系 / モルティエレラ / 糸状菌 / ヒドロキシ脂肪酸 / スーパーオキシドジスムターゼ / 還元反応 / 有機酸 / 嫌気反応 / 共役化反応 / 飽和化反応 / 嫌気性微生物 / 嫌気性菌 / 共役脂肪 / Lacctobacillus / Eubacterium / 嫌気性細菌 / Lactobacillus / キラル化合物 / 不斉還元 / 旧黄色酵素 / カルボニル還元酵素 / Chiral compound / Asymmetric reduction / Old yellow enzyme / Carbonyl reductase / バイオ不斉還元 / アミノアルコート脱水素酵素 / ランダム変異 / Agrobacterium / Rhodococcus / アミノアルコール脱水素酵素 / Hansenula / chiral compound / carbonyl reductase / bioreduction / aminoalcohol dehydrogenase / random mutation / 電子伝達 / インジゴ / 藍染 / 藍染め / 複合微生物系 / 耐熱性 / アルカリ耐性 / 酸化還元

研究課題研究成果共同研究者

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2019年11月12日

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櫻谷英治

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子実体形成誘導技術を応用したマツタケ栽培技術の開発（阪本鷹行）

食用コオロギの機能性検証と生産システムの社会実装

高大連携による県内希少植物イシマササユリの香気成分の解析および研究開発の推進

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櫻谷 英治

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櫻谷英治