Construction of efficient conversion system from L-arabinose to ethanol by Saccharomyces cerevisiae

2008 Fiscal Year Final Research Report

• Project/Area Number: 18760592
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Biofunction/Bioprocess
• Research Institution: Kyoto University
• Principal Investigator: WATANABE Seiya Kyoto University, 生存基盤科学研究ユニット, 特定研究員 (90379032)
• Project Period (FY): 2006 – 2008
• Keywords: バイオマス / エタノール / サッカロミセス酵母 / L-アラビノース / 生物発酵

Research Abstract

木質系バイオマスに含まれる五炭糖であるL-アラビノースのサッカロミセス酵母によるエタノール発酵を目指して、窒素固定細菌の一種から既知のものとは全く異なるL-アラビノース代謝経路の生化学的・分子生物学的・構造生物学的解析を行った。また、リグノセルロースバイオマス成分の1つであるL-ラムノースにおいても、本経路と類似の新規経路が酵母および細菌の一部に存在することを初めて発見した。

Research Products

• [Journal Article] Novel modified version of non-phosphorylated sugar metabolism: an alternative L-rhamnose pathway of Sphingomonas sp (2009)
  • Author(s): Watanabe, S. and Makino, K
  • Journal Title: FEBS Journal 276 (6) Pages: 1554-1567
• [Journal Article] Eukaryotic and bacterial gene clusters related to an alternative pathway of non-phosphorylated L-rhamnose metabolism (2008)
  • Author(s): Watanabe, S., Saimura, M. and Makino, K
  • Journal Title: Journal of Biological Chemistry 283 Pages: 20372-20382
• [Journal Article] Gene cloning of cold-adapted isocitrate lyase from a psychrophilic bacterium, Colwellia psychrerythraea, and analysis of amino acid residues involved in cold adaptation of this enzyme (2008)
  • Author(s): Sato, Y., Watanabe, S., Yamaoka, N. and Takada, Y
  • Journal Title: Extremophiles 12 (1) Pages: 107-117
• [Journal Article] Bioethanol production from xylose by recombinant Saccharomyces cerevisiae expressing xylose reductase, NADP+-dependent xylitol dehydrogenase, and xylulokinase (2008)
  • Author(s): Matsushika, A., Watanabe, S., Kodaki, T., Makino, K. and Sawayama, S
  • Journal Title: Journal of Bioscience and Bioengineering 105 (3) Pages: 296-299
• [Journal Article] Expression of protein engineered NADP+-dependent xylitol dehydrogenase increases ethanol production from xylose in recombinant Saccharomyces cerevisiae (2008)
  • Author(s): Matsushika, A., Watanabe, S., Kodaki, T., Makino, K., Inoue, H., Murakami, K. and Sawayama, S
  • Journal Title: Applied Microbiology and Biotechnology 81 (2) Pages: 243-255
• [Journal Article] Ethanol production from xylose by recombinant Saccharomyces cerevisiae expressing protein engineered NADP+-dependent xylitol dehydrogenase (2007)
  • Author(s): Watanabe, S., Saleh, A.A., Pack, S.P., Annaluru, N., Kodaki, T. and Makino, K
  • Journal Title: Journal of Biotechnology 130 (3) Pages: 316-319
• [Journal Article] Protein engineering of xylose reductase from Pichia stipitis for improved NADH-specificity and the efficient ethanol production from xylose in recombinant Saccharomyces cerevisiae (2007)
  • Author(s): Watanabe, S., Saleh, A.A., Pack, S.P., Annaluru, N., Kodaki, T. and Makino, K
  • Journal Title: Microbiology 153 (9) Pages: 3045-3055
• [Journal Article] The Positive effect of the decreased NADPH-preferring activity of xylose reductase from Pichia stipitis on the ethanol production using xylose-fermenting recombinant Saccharomyces cerevisiae (2007)
  • Author(s): Watanabe, S., Pack, S.P., Saleh, A.A., Pack, S.P., Annaluru, N., Kodaki, T. and Makino, K
  • Journal Title: Bioscience, Biotechnology and Biochemistry 71 (5) Pages: 1365-1369
• [Journal Article] Thermostabilization of Pichia stipitis xylitol dehydrogenase by mutation of structural zinc binding loop (2007)
  • Author(s): Annaluru, N., Watanabe, S., Pack, S.P., Saleh, A.A., Kodaki, T., and Makino, K
  • Journal Title: Journal of Biotechnology 129 (4) Pages: 717-722
• [Journal Article] Preliminary crystallographic analysis of L-2-keto-3-deoxyarabonate dehydratase involved in an alternative bacterial pathway of L-arabinose metabolism (2007)
  • Author(s): Shimada, N., Mikami, B., Watanabe, S., Makino, K
  • Journal Title: Acta Crystallographica F63 (5) Pages: 393-395
• [Journal Article] Base sequence- and Tm-dependent DNA oligomer separation by open tubular capillary columns carrying complementary DNA oligomers as probes (2007)
  • Author(s): Devarayapalli, K.C., Pack, S.P., Kamisetty, N.K., Nonogawa, M., Watanabe, S., Kodaki, T. and Makino, K
  • Journal Title: Anaytical and Bioanalytical Chemistry 388 (4) Pages: 919-928
• [Journal Article] Efficient preparation of amine-modified oligodeoxynucleotide using modified H-phosphonate chemistry for DNA microarray fabrication (2007)
  • Author(s): Kamisetty, N.K., Pack, S.P., Nonogawa, M., Devarayapalli, K.C., Watanabe, S., Kodaki, T. and Makino, K
  • Journal Title: Anaytical and Bioanalytical Chemistry 387 (6) Pages: 2027-2035
• [Journal Article] Cloning, expression and characterization of bacterial L-arabinose 1-dehydrogenase involved in an alternative pathway of L-arabinose metabolism (2006)
  • Author(s): Watanabe, S., Kodaki, T. and Makino, K
  • Journal Title: Journal of Biological Chemistry 281 (5) Pages: 2612-2623
• [Journal Article] A novel α-ketoglutaric semialdehyde dehydrogenase : evolutionary insight into an alternative pathway of bacterial L-arabinose metabolism (2006)
  • Author(s): Watanabe, S., Kodaki, T. and Makino, K
  • Journal Title: Journal of Biological Chemistry 281 (39) Pages: 28876-28888
• [Journal Article] Identification and characterization of L-arabonate dehydratase, L-2-keto-3-deoxyarabonate dehydratase and L-arabinolactonase involved in an alternative pathway of L-arabinose metabolism : novel evolutionary insight into sugar metabolism (2006)
  • Author(s): Watanabe, S., Shimada, N., Tajima, K., Kodaki, T. and Makino, K
  • Journal Title: Journal of Biological Chemistry 281 (44) Pages: 33521-33536
• [Journal Article] Construction of various mutants of xylose metabolizing enzymes for efficient conversion of biomass to ethanol (2006)
  • Author(s): Saleh, A.A., Watanabe, S., Annaluru, N., Kodaki, T. and Makino, K
  • Journal Title: Nucleic Acids Symposium Series 50 Pages: 279-280
• [Journal Article] Site-directed mutagenesis of a yeast gene for improvement of enzyme thermostability (2006)
  • Author(s): Annaluru, N., Watanabe, S., Saleh, A.A., Kodaki, T., and Makino, K
  • Journal Title: Nucleic Acids Symposium Series 50 Pages: 281-282
• [Journal Article] Metabolic fate of L-lactaldehyde derived from an alternative L-rhamnose pathway
  • Author(s): Watanabe, S., Piyanart, S. and Makino, K
  • Journal Title: FEBS Journal 275 (20) Pages: 5139-5149
• [Journal Article] α-Ketoglutaric semialdehyde dehydrogenase isozymes involved in metabolic pathways of D-glucarate, D-galactarate and hydroxy-L-proline : molecular and metabolic convergent evolution
  • Author(s): Watanabe, S., Yamada, M., Ohtsu, I. and Makino, K
  • Journal Title: Journal of Biological Chemistry 282 (9) Pages: 6685-6695
• [Presentation] タンパク質工学を用いたキシロース発酵性サッカロミセス酵母の育種 (2008)
  • Author(s): 渡辺誠也
  • Organizer: 第60回日本生物工学会大会
  • Place of Presentation: 東北学院大学(仙台)
  • Year and Date: 20080827-29
• [Presentation] Bioethanol production from xylose by recombinant Saccharomyces cerevisiae expressing xylose reductase, NADP+-dependent xylitol dehydrogenase, and xylulokinase (2008)
  • Author(s): Matsushika, A., Watanabe, S., Kodaki, T., Makino, K. and Sawayama, S
  • Organizer: 30th Symposium on Biotechnology for Fuels and Chemicals
  • Place of Presentation: ニューオリンズ
  • Year and Date: 20080504-07
• [Presentation] 改変型キシリトール脱水素酵素を導入した酵母を用いたキシロースからのエタノール生産 (2008)
  • Author(s): 松鹿昭則、渡辺誠也、小瀧努、牧野圭祐、井上宏之、村上克治、澤山茂樹
  • Organizer: 2008年度農芸化学会年会
  • Place of Presentation: 名城大学(名古屋)
  • Year and Date: 20080326-29
• [Presentation] 新規アラビノース代謝経路に関与するL-2-ケト-3-デオキシアラボネート脱水酵素の結晶構造解析 (2007)
  • Author(s): 嶋田直子、三上文三、渡辺誠也、小瀧努、牧野圭祐
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
  • Place of Presentation: パシフィコ横浜(横浜)
  • Year and Date: 20071211-15
• [Presentation] 新規L-アラビノース代謝経路に関与するL-2-keto-3-deoxyarabonate脱水酵素のX線構造解析 (2007)
  • Author(s): 嶋田直子、三上文三、渡辺誠也、小瀧努、牧野圭祐
  • Organizer: 日本結晶学会2007年度年会
  • Place of Presentation: 東京
  • Year and Date: 20071201-02
• [Presentation] 改変型酵素を導入した酵母によるキシロースからのエタノール生産 (2007)
  • Author(s): 松鹿昭則、渡辺誠也、小瀧努、牧野圭祐、井上宏之、村上克治、澤山茂樹
  • Organizer: 第59回日本生物工学会大会
  • Place of Presentation: 広島大学(広島)
  • Year and Date: 20070925-27
• [Presentation] 細菌由来の新規なL-アラビノース代謝経路 (2007)
  • Author(s): 渡辺誠也、牧野圭祐
  • Organizer: 分子生物学会 第7回春季シンポジウム
  • Place of Presentation: 淡路島
  • Year and Date: 20070422-24
• [Presentation] 脱水素酵素の補酵素特異性改変と安定化戦略 (2007)
  • Author(s): 渡辺誠也、小瀧努、牧野圭祐
  • Organizer: 酵素化学ミニシンポジウム
  • Place of Presentation: 京都大学(京都)
  • Year and Date: 2007-09-10
• [Presentation] Highly Efficient Bioethanol Fermentation from Woody Biomass (2007)
  • Author(s): Watanabe, S., Kodaki, T. and Makino, K
  • Organizer: Core university program joint seminar between Seoul national university and Kyoto university ~Research and development for environmentally clean renewable energy production system~
  • Place of Presentation: 京都大学(京都)
  • Year and Date: 2007-07-20
• [Presentation] 酵母の糖代謝経路の新展開-バイオエタノール生産への活用から新規代謝経路の発見まで- (2007)
  • Author(s): 渡辺誠也
  • Organizer: 第10回真核微生物交流会
  • Place of Presentation: 酒類総合研究所(広島)
  • Year and Date: 2007-06-22
• [Presentation] Improvement of thermostability of Pichia stipitis xylitol dehydrogenase by protein engineering (2006)
  • Author(s): Annaluru, N., Watanabe, S., Saleh, A. A., Kodaki, T. and Makino, K
  • Organizer: 第33回核酸化学シンポジウム
  • Place of Presentation: 大阪市
  • Year and Date: 20061120-22
• [Presentation] New approach for conversion of biomass to ethanol by recombinant Saccharomyces cerevisiae (2006)
  • Author(s): Saleh, A. A., Watanabe, S., Annaluru, N., Kodaki, T. and Makino, K
  • Organizer: 第33回核酸化学シンポジウム
  • Place of Presentation: 大阪市
  • Year and Date: 20061120-22
• [Presentation] Development of highly efficient fermentation process for bioethanol from biomass based on novel protein engineering approach (2006)
  • Author(s): Watanabe, S., Saleh, A. A., Annaluru, N., Shimada, N., Kodaki, T. and Makino, K
  • Organizer: 5大学連携21COE合同国際シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 20061005-08
• [Presentation] A novel bacterial L-arabinose pathway: cloning, expression and characterization of the metabolic genes (2006)
  • Author(s): Watanabe, S., Shimada, N., Kodaki, T. and Makino, K
  • Organizer: The 3rd International Symposium on Sustainable Energy System
  • Place of Presentation: 京都大学
  • Year and Date: 20060930-1001
• [Presentation] Construction of thermostable xylitol dehydrogenase, a key enzyme for efficient bioethanol production from xylose (2006)
  • Author(s): Annaluru, N., Watanabe, S., Saleh, A. A., Kodaki, T. and Makino, K
  • Organizer: The 3rd International Symposium on Sustainable Energy System
  • Place of Presentation: 京都大学
  • Year and Date: 20060930-1001
• [Presentation] New approach for conversion of biomass to ethanol by recombinant Saccharomyces cerevisiae (2006)
  • Author(s): Saleh, A. A., Watanabe, S., Annaluru, N., Kodaki, T. and Makino, K
  • Organizer: The 3rd International Symposium on Sustainable Energy System
  • Place of Presentation: 京都大学
  • Year and Date: 20060930-1001
• [Presentation] A novel L-arabonate dehydratase involved in an alternative bacterial pathway of L-arabinose metabolism (2006)
  • Author(s): Shimada, N., Watanabe, S., Shimada, N., Kodaki, T. and Makino, K
  • Organizer: The 3rd International Symposium on Sustainable Energy System
  • Place of Presentation: 京都大学
  • Year and Date: 20060930-1001
• [Presentation] Bioethanol from woody biomass by supercritical water technology -highly efficient ?fermentation process for bioethanol from woody biomass- (2006)
  • Author(s): Makino, K., Saleh, A. A., Watanabe, S., Annaluru, N., and Kodaki, T
  • Organizer: The 3rd International Symposium on Sustainable Energy System
  • Place of Presentation: 京都大学
  • Year and Date: 20060930-1001
• [Presentation] Improvement of thermostability of Pichia stipitis xylitol dehydrogenase by protein engineering (2006)
  • Author(s): Annaluru, N., Watanabe, S., Saleh, A. A., Kodaki, T. and Makino, K
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: 京都
  • Year and Date: 20060618-23
• [Presentation] New approach for conversion of biomass to ethanol by recombinant Saccharomyces cerevisiae (2006)
  • Author(s): Saleh, A. A., Watanabe, S., Annaluru, N., Kodaki, T. and Makino, K
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: 京都
  • Year and Date: 20060618-23
• [Presentation] Crystallization and preliminary X-ray diffraction studies of xylitol dehydrogenase, a key enzyme for bioethanol production using yeas (2006)
  • Author(s): Matsui, J., Watanabe, S., Kodaki, T. and Makino, K
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: 京都
  • Year and Date: 20060618-23
• [Presentation] A novel bacterial L-arabinose pathway -cloning, expression and characterization of the metabolic genes- (2006)
  • Author(s): Watanabe, S., Kodaki, T. and Makino, K
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: 京都
  • Year and Date: 20060618-2
• [Presentation] 五炭糖代謝に関わる酵素のタンパク質工学による機能改変 (2006)
  • Author(s): 依田香子、渡辺誠也、小瀧努、牧野圭祐
  • Organizer: 日本化学会第86回春季大会
  • Place of Presentation: 千葉
  • Year and Date: 20060327-30
• [Book] 産業酵素の応用技術と最新動向, (第24章 サッカロミセス酵母による効率的キシロース発酵を目指したキシリトール脱水素酵素の機能変換) (2009)
  • Author(s): 渡辺誠也
  • Publisher: シーエムシー出版
• [Book] 化学と教育56 (12), (木質系バイオマスからのエタノール生産のための微生物育種戦略) (2008)
  • Author(s): 渡辺誠也
• [Book] 生物工学会誌 第11号, (-さまざまな微生物による燃料用エタノール製造法の進展-タンパク質工学を用いたキシロース発酵性サッカロミセス酵母の育種) (2008)
  • Author(s): 渡辺誠也
  • Total Pages: 19
• [Book] 生化学 79 (11), (微生物の糖代謝経路に見られる新規な進化学的関係) (2007)
  • Author(s): 渡辺誠也、牧野圭祐
  • Total Pages: 1059-1064
• [Book] バイオサイエンスとインダストリー 65 (12), (タンパク質工学によるキシロース発酵酵母作出の新戦略) (2007)
  • Author(s): 渡辺誠也、牧野圭祐
  • Total Pages: 18-20
• [Book] 化学フロンティア17, (新エネルギー最前線-環境調和型エネルギーシステムの構築を目指して-バイオエタノール-遺伝子工学による発酵技術の改変-) (2006)
  • Author(s): 渡辺誠也、小瀧努、牧野圭祐
  • Total Pages: 107-114
• [Patent(Industrial Property Rights)] リンカー化合物、プローブ及びプローブ固定化担体 (2006)
  • Inventor(s): 牧野圭祐、小瀧努、渡辺誠也、スンピル パック、野々川満、和田啓男、吉田安子、山田和成
  • Industrial Property Rights Holder: 京都大学、日本碍子、信和化工 (株)
  • Industrial Property Number: PCT/JP2006/304641
  • Filing Date: 2006-03-09
  • Overseas
• [Patent(Industrial Property Rights)] L-アラビノースをα-ケトグルタル酸に変換する方法、この方法に使用する酵素、該酵素をコードする遺伝子、該遺伝子を含む組換え微生物 (2005)
  • Inventor(s): 牧野圭祐、小瀧努、渡辺誠也、和田啓男
  • Industrial Property Rights Holder: 京都大学、信和化工(株)
  • Industrial Property Number: 特願2005-303253
  • Filing Date: 2005-10-18