1996 Fiscal Year Annual Research Report

微生物より単離した有用酵素を用いた不斉変換反応

Research Project

Project/Area Number	07454194
Research Institution	KYOTO UNIVERSITY
Principal Investigator	大野惇吉京都大学, 化学研究所, 教授 (70027077)
Keywords	不斉還元 / パン酵母 / 炭素-炭素二重結合還元酵素 / 不斉合成 / ケトエステル還元酵素 / Geotrichum candidum
Research Abstract	有機合成化学的に有用な不斉触媒として新しい酵素を微生物中から探索,単離.,精製し,様々な化合物の立体,官能基,位置選択的な変換反応を確立するのが申請者の計画である.微生物の代わりに精製した酵素を生体触媒として用いる最大の利点は,共存する酵素による副反応が防げるという利点である.微生物を用いた反応では副反応によって満足する結果が得られていない様な反応に的を絞って酵素の探索,単離,精製を行った. まず微生物として最も汎用的なパン酵母を選び,ケトエステル還元酵素の単離精製を行ったところ,α-ケトエステル還元酵素が七種類とβ-ケトエステル還元酵素四種類を単離する事ができた.それぞれの酵素を使い分けることで様々な構造を有するαおよびβ-ヒドロキシエステルのR体とS体の作り分けが可能になった.またこれらの酵素の中には,全くのラセミ化合物から一段階の反応で三つの不斉炭素を立体選択的に導入する酵素があることも見出した. 次にα,β-不飽和ケトンの炭素-炭素二重結合を選択的に還元する酵素をパン酵母中から単離精製する事に成功した.この酵素による還元反応を系統的に検討したところ,立体選択性は基質の構造に大きく依存する事を見出した.そして,種々の4-aryl-3-methyl-3-buten-2-oneの官能基および立体選択的な還元反応に成功し,高立体選択的な光学活性カルボニル化合物の合成法を確立した.また,本酵素はニトロオレフィンの炭素-炭素二重結合も還元できることを見出し,光学活性なニトロ化合物の合成法を確立した. 次に,培養および集菌が容易なチチカビの一種であるGeotrichum candidumからケトエステル還元酵素の単離を行ったところ,ethyl2-methyl-3-oxobutanoateに還元活性のある酵素を三種類単離精製することに成功した.これらの酵素を使い分けることで(2R,3S)体と(2S,3S)体のヒドロキシエステルを作り分けることが可能になった.

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] Yasushi Kawai: "Stereochemical Control in Microbial Reduction XXV.Additives Controlling Diastereoselectivity in a Microbial Reduction of Ethyl 2-Methyl-3-oxobutanoate." Bulletin of the Chemical Society of Japan. 68. 285-288 (1995)
[Publications] Atsuyoshi Ohono: "NAD(P)^+-NAD(P)H Models.84.Stereochemistry Controlled by Electronic Effect from a Sulfinyl Group." Heteroatom Chemistry. 6. 51-56 (1995)
[Publications] Kaoru Nakamura: "Improvement of Enantioselectivity of Microbial Reduction by Using Organic Solvent Redox Coupler System." Tetrahedron Letters. 36. 265-266 (1995)
[Publications] Yasushi Kawai: "Introduction of Plural Asymmetric Centers by a β-Keto Ester Reductase from Bakers′ Yeast." Tetrahedron Letters. 36. 591-592 (1995)
[Publications] Kaoru Nakamura: "Mechanistic Study for Stereochemical Control of Microbial Reduction of α-Keto Esters in an Organic Solvent." Tetrahedron. 51. 687-694 (1995)
[Publications] Kaoru Nakamura: "Microbial Deracemization of 1-Arylethanol." Tetrahedron Letters. 36. 6263-6266 (1995)
[Publications] Atsuyoshi Ohono: "NAD(P)^+-NAD(P)H Models.87.Nonsteric Stereochemistry Controlled by a Carbonyl Dipole" Bulletin of the Chemical Society of Japan. 69. 1679-1685 (1996)
[Publications] Masamichi Kinoshita: "Factors Influencing Enantioselectivity of Lipase-Catalyzed Hydrolysis" Tetrahedron. 52. 5397-5406 (1996)
[Publications] Yasushi Kawai: "Stereochemical Control in Microbial Reduction XXVIII.Asymmetric Reduction of α,β-Unsaturated Ketones with Bakers′ Yeast" Bulletin of the Chemical Society of Japan. 69. 2633-2638 (1996)
[Publications] Kaoru Nakamura: "Different Stereochemistry for the Reduction of Trifluoromethyl Ketones and Methyl Ketones Catalyzed by Alcohol Dehydrogenase from Geotrichum" Tetrahedron Letters. 37. 5727-5730 (1996)
[Publications] Kaoru Nakamura: "Asymmetric Synthesis of(S)-Arylalkanols by Microbial Reduction" Tetrahedron : Asymmetry. 7. 3021-3024 (1996)
[Publications] Atsuyoshi Ohno: "Atropisomeric Flavoenzyme Models with a Modified Pyridine Ring : Synthesis,Physical Properties,and Stereochemistry in the Reactions with NAD(P)H Analogs" The Journal of Organic Chemistry. 61. 9344-9355 (1996)
[Publications] Kaoru Nakamura: "Structure of Solvent Affects Enantioselectivity of Lipase-catalyzed Transesterification." Tetrahedron. 51. 8799-8088 (1995)
[Publications] Yasushi Kawai: "Asymmetric Reduction of α,β-Unsaturated Ketones with Bakers′ Yeast." Tetrahedron : Asymmetry. 6. 2143-2144 (1995)
[Publications] Kaoru Nakamura: "Lipase-Catalyzed Transesterification of Aryl-Substituted Alkanols in an Organic Solvent" Bulletin of the Chemical Society of Japan. 69. 1079-1085 (1996)
[Publications] Atsuyoshi Ohno: "NAD(P)^+-NAD(P)H Models.86.Nonsteric Stereochemistry in Hydride-Transfer to Sulfinylpyridinium Ion" Bulletin of the Chemical Society of Japan. 69. 1093-1098 (1996)
[Publications] Kaoru Nakamura: "Arymmetric Redyction of Ketones by Acetone Powder of Geotrichum candidum" Tetrahedron Letters. 37. 1629-1632 (1996)
[Publications] Kaoru Nakamura: "Enantio-and Regioselective Reduction of α-Diketones by Bakers′ Yeast" Tetrahedron : Asymmetry. 7. 409-412 (1996)

1996 Fiscal Year Annual Research Report

微生物より単離した有用酵素を用いた不斉変換反応

Principal Investigator

大野 惇吉 京都大学, 化学研究所, 教授 (70027077)

Research Products

[Publications] Yasushi Kawai: "Stereochemical Control in Microbial Reduction XXV.Additives Controlling Diastereoselectivity in a Microbial Reduction of Ethyl 2-Methyl-3-oxobutanoate." Bulletin of the Chemical Society of Japan. 68. 285-288 (1995)

[Publications] Atsuyoshi Ohono: "NAD(P)^+-NAD(P)H Models.84.Stereochemistry Controlled by Electronic Effect from a Sulfinyl Group." Heteroatom Chemistry. 6. 51-56 (1995)

[Publications] Kaoru Nakamura: "Improvement of Enantioselectivity of Microbial Reduction by Using Organic Solvent Redox Coupler System." Tetrahedron Letters. 36. 265-266 (1995)

[Publications] Yasushi Kawai: "Introduction of Plural Asymmetric Centers by a β-Keto Ester Reductase from Bakers′ Yeast." Tetrahedron Letters. 36. 591-592 (1995)

[Publications] Kaoru Nakamura: "Mechanistic Study for Stereochemical Control of Microbial Reduction of α-Keto Esters in an Organic Solvent." Tetrahedron. 51. 687-694 (1995)

[Publications] Kaoru Nakamura: "Microbial Deracemization of 1-Arylethanol." Tetrahedron Letters. 36. 6263-6266 (1995)

[Publications] Atsuyoshi Ohono: "NAD(P)^+-NAD(P)H Models.87.Nonsteric Stereochemistry Controlled by a Carbonyl Dipole" Bulletin of the Chemical Society of Japan. 69. 1679-1685 (1996)

[Publications] Masamichi Kinoshita: "Factors Influencing Enantioselectivity of Lipase-Catalyzed Hydrolysis" Tetrahedron. 52. 5397-5406 (1996)

[Publications] Yasushi Kawai: "Stereochemical Control in Microbial Reduction XXVIII.Asymmetric Reduction of α,β-Unsaturated Ketones with Bakers′ Yeast" Bulletin of the Chemical Society of Japan. 69. 2633-2638 (1996)

[Publications] Kaoru Nakamura: "Different Stereochemistry for the Reduction of Trifluoromethyl Ketones and Methyl Ketones Catalyzed by Alcohol Dehydrogenase from Geotrichum" Tetrahedron Letters. 37. 5727-5730 (1996)

[Publications] Kaoru Nakamura: "Asymmetric Synthesis of(S)-Arylalkanols by Microbial Reduction" Tetrahedron : Asymmetry. 7. 3021-3024 (1996)

[Publications] Atsuyoshi Ohno: "Atropisomeric Flavoenzyme Models with a Modified Pyridine Ring : Synthesis,Physical Properties,and Stereochemistry in the Reactions with NAD(P)H Analogs" The Journal of Organic Chemistry. 61. 9344-9355 (1996)

[Publications] Kaoru Nakamura: "Structure of Solvent Affects Enantioselectivity of Lipase-catalyzed Transesterification." Tetrahedron. 51. 8799-8088 (1995)

[Publications] Yasushi Kawai: "Asymmetric Reduction of α,β-Unsaturated Ketones with Bakers′ Yeast." Tetrahedron : Asymmetry. 6. 2143-2144 (1995)

[Publications] Kaoru Nakamura: "Lipase-Catalyzed Transesterification of Aryl-Substituted Alkanols in an Organic Solvent" Bulletin of the Chemical Society of Japan. 69. 1079-1085 (1996)

[Publications] Atsuyoshi Ohno: "NAD(P)^+-NAD(P)H Models.86.Nonsteric Stereochemistry in Hydride-Transfer to Sulfinylpyridinium Ion" Bulletin of the Chemical Society of Japan. 69. 1093-1098 (1996)

[Publications] Kaoru Nakamura: "Arymmetric Redyction of Ketones by Acetone Powder of Geotrichum candidum" Tetrahedron Letters. 37. 1629-1632 (1996)

[Publications] Kaoru Nakamura: "Enantio-and Regioselective Reduction of α-Diketones by Bakers′ Yeast" Tetrahedron : Asymmetry. 7. 409-412 (1996)

大野惇吉京都大学, 化学研究所, 教授 (70027077)