Comprehensive screening of novel microbial enzymes toward on-demand production of hydroxy amino acids

2010 Fiscal Year Final Research Report

• Project/Area Number: 21780070
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied microbiology
• Research Institution: Kyoto University
• Principal Investigator: HIBI Makoto Kyoto University, 大学院・農学研究科, 助教 (30432347)
• Project Period (FY): 2009 – 2010
• Keywords: 水酸化アミノ酸 / 微生物スクリーニング / ジオキシゲナーゼ / 酵素触媒

Research Abstract

Hydroxy amino acids are promising compounds for the use to drugs and foods, because some of them possess useful physiological activities. Then efficient production method for hydroxy amino acids is desired. In this research, four dioxygenases obtained from microorganisms were found to catalyze hydroxylation of amino acids, and their substrate specificities and chemical structure of the products were determined. The hydroxylating activities of these dioxygenases were novel types of reactions, and all of their reaction was highly stereospecific. Thus these dioxygenases were useful for the biocatalysts for the production of chiral hydroxy amino acids.

Research Products

• [Journal Article] A novel L-isoleucine metabolism i n Bacillus thuringiensis generating (2S, 3R,4S)-4-hydroxyisoleucine (2011)
  • Author(s): Ogawa J, Kodera T, Smirnov SV, Hibi M, Samsonova NN, Koyama R, Yamanaka H, Mano J, Kawashima T, Yokozeki K, Shimiz u S.
  • Journal Title: a potential insulinotropic and anti-obesity amino acid, Applied Microbiology and Biotechn olog 89 Pages: 1929-1938
  • Peer Reviewed
• [Journal Article] Metabolic engineering of Escherich ia coli to produce (2S,3R,4S)-4-hydroxy isoleucine (2010)
  • Author(s): Smirnov SV, Kodera T , Samsonova NN , Kotlyarova VA, Rushkevich NY, Kivero AD, Sokolov PM, Hibi M, Ogawa J, Shimizu S.
  • Journal Title: Applied Microbiology and Bi otechnology 88 Pages: 719-726
  • Peer Reviewed
• [Journal Article] A novel l-isoleuc ine hydroxylating enzyme, l-isoleucine dioxygenase from Bacillus thuringiensis, produces (2S,3R,4S)-4-hydroxyisoleucine (2009)
  • Author(s): Kodera T, Smirnov SV, Samsonova NN, Kozlov YI, Koyama R, Hibi M, Ogawa J, Y okozeki K, Shimizu S
  • Journal Title: Biochemical and Biophysical Research Communications. 390 Pages: 506-510
  • Peer Reviewed
• [Presentation] Bacillus thuringiensis 2e2株由来L-isoleucine dioxygenaseによる有用アミノ酸の立体選択的合成 (2011)
  • Author(s): 日比慎、河嶋隆志、清水昌、横関健三、小川順
  • Organizer: 日本農芸化学会2011年度大会
  • Place of Presentation: 京都
  • Year and Date: 2011-03-26
• [Presentation] Burkholderia ambifariaAMMD株由来新規dioxygenaseの機能解析及び反応生成物の同定 (2011)
  • Author(s): 河嶋隆志, 日比慎, 横関健三, 清水昌, 小川順
  • Organizer: 日本農芸化学会2011年度大会
  • Place of Presentation: 京都
  • Year and Date: 2011-03-26
• [Presentation] 微生物由来新規アミノ酸水酸化酵素の基質特異性解析及び生成物の同定 (2010)
  • Author(s): 河嶋隆志, 日比慎, 横関健三, 清水昌, 小川順
  • Organizer: 第62回日本生物工学会
  • Place of Presentation: 宮崎
  • Year and Date: 2010-10-28
• [Presentation] Application of microbial enzymes in the production of functional hydroxy amino acids (2010)
  • Author(s): Makoto Hibi, Jun Ogawa, Kenzo Yokozeki, Sakayu Shimizu
  • Organizer: The 12th Japanese-Swiss Meeting on Biotechnology and Bioprocess Development
  • Place of Presentation: 富山
  • Year and Date: 2010-10-11
• [Presentation] 微生物由来新規アミノ酸水酸化酵素の探索及び基質特異性解析 (2010)
  • Author(s): 河嶋隆志, 日比慎, 小川順, 横関健三, 清水昌
  • Organizer: 日本農芸化学会2010年度大会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-28
• [Presentation] Bacillus属細菌における新規L-イソロイシン代謝経路とその生理的意義 (2009)
  • Author(s): 日比慎, 小川順, 横関健三, 清水昌
  • Organizer: 2009年度日本農芸化学会関西・中四国・西日本支部、日本栄養・食糧学会九州・沖縄支部および日本食品科学工学会西日本支部合同沖縄大会
  • Place of Presentation: 沖縄
  • Year and Date: 2009-10-31
• [Presentation] Bacillus属細菌由来4-ヒドロキシイソロイシン脱水素酵素の機能解析 (2009)
  • Author(s): 日比慎, 河嶋隆志, 小川順, 横関健三, 清水昌
  • Organizer: 2009年度日本農芸化学会関西・中四国・西日本支部、日本栄養・食糧学会九州・沖縄支部および日本食品科学工学会西日本支部合同沖縄大会
  • Place of Presentation: 沖縄
  • Year and Date: 2009-10-31
• [Presentation] Enzymatic synthesis of 4-hydroxy isoleucine with novel dioxygenase (2009)
  • Author(s): Makoto Hibi, Jun Ogawa, Tomohiro Kodera, Sergey V.Smirnov, Kenzo Yokozeki, Sakayu Shimizu
  • Organizer: 15th German-Japanese Workshop on Enzyme Technology, Rostock
  • Place of Presentation: Germany
  • Year and Date: 2009-09-26
• [Presentation] Enzymatic Synthesis of 4-Hydroxy isoleucine with Novel Dioxygenase, Enzyme (2009)
  • Author(s): M.Hibi, J.Ogawa, T.Kodera, S.V.Smirnov, K. Yokozeki, S.Shimizu
  • Organizer: Engineering XX, Groningen
  • Place of Presentation: Netherlands
  • Year and Date: 2009-09-23
• [Patent(Industrial Property Rights)] Method for producing hydroxylated amino acids (2011)
  • Inventor(s): Smirnov, S.V., Samsonova, N.N., Kotliarova, V.A., Rushkevich, N.Y., Fedorina, E.A., Sokolov, P.M., Kolokolova, A.V., Ogawa, J., Hibi, M., Shimizu, S., Imabayashi, Y., Suzuki, S., Sugiyama, M.
  • Industrial Property Rights Holder: Ajinomoto Co., Inc.
  • Industrial Property Number: PTC特許,PT2011021717
  • Filing Date: 2011-02-24
  • Overseas