New Function of Halohydrin Epoxidase and Its Application of the Synthesis of Chiral Building Block.

卤代醇环氧化酶的新功能及其在手性构件合成中的应用。

• 批准号: 05660088
• 负责人: NAGASAWA Toru
• 金额: $ 1.41万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660088/
• 关键词: Halohydrin Hydrogen Halide-lyase; Epoxide Hydrolase; Chiral Glyceral Devivative; (R)-3-Chloro-1,2-propanediol; Prochiral; Epichlorohydrin; halohydrinhydrogen-halide-lyas; epoxide hydrolase; (R)-3クロロ-1,2-プロパンジオール; キラルソース; ハロヒドリン; エピフロルヒドリン; プロキ

We investigated the microbial production of (R)-3-chloro-1,2-propanediol (MCP) from prochiral 1,3-dichloro-2-propanol (DCP) and enzymes involved in the transformation of DCP into (R)-MCP.Microorganisms which utilize MCP or epichlorohydrin (ECH) as a sole carbon source were isolated from soil samples by means of an enrichment culture technique. The production of (R)-MCP from DCP was examined with one of these strains, Corynebacterium sp.strain N-1074. The optical purity of (R)-MCP formed was improved by keeping the level of DCP in the reaction mixture at a low concentration. (R)-MCP was obtained from 77.5 mM DCP with a 97.3% molar conversion yield and an 83.8% e.e.of its optical purity by periodic feeding of the substrate. During the course of the transformation of DCP into (R)-MCP with the cell extract of Corynebacterium sp.strain N-1074, ECH was transiently formed. The cell extract was fractionated into two DCP-dechlorinating activities and two ECH-hydrolyzing activities. Each enzyme was purified and characterized. DCP-dechlorinating activities and two ECH-hydrolyzing activities were proved to be halohydrin hydrogen halide-lyase and epoxide hydrolase, respectively. Each one of them exhibited the sentioselectivity for the conversion of DCP into ECH and ECH into MCP.We also investigated the application of the dehalogenating enzymes catalyzing the interconversion of halohydrins into the corresponding epoxides to the synthesis of beta-hydroxynitriles.

采用富集培养技术，从土壤样品中分离出以1，3-二氯-2-丙醇（DCP）和环氧氯丙烷（ECH）为唯一碳源的微生物，研究了DCP转化为（R）-MCP的酶系及微生物转化产（R）-3-氯-1，2-丙二醇（MCP）的过程。用这些菌株中的一种，棒状杆菌属菌株N-1074，检测从DCP产生（R）-MCP。通过将反应混合物中DCP的水平保持在低浓度来提高所形成的（R）-MCP的光学纯度。（R）-MCP由77.5mM DCP通过周期性进料底物以97.3%摩尔转化率和83. 8% e.e.的光学纯度获得。在用棒状杆菌N-1074菌株的细胞提取物将DCP转化为（R）-MCP的过程中，瞬时形成ECH。细胞提取物分为两个DCP-脱氯活性和两个ECH-水解活性。对每种酶进行纯化和表征。DCP脱氯活性和ECH水解活性分别为卤代醇氢裂解酶和环氧化物水解酶。它们对DCP转化为ECH和ECH转化为MCP均表现出一定的选择性。我们还研究了催化卤代醇转化为相应环氧化物的脱卤酶在β-羟基腈合成中的应用。

项目成果

Tetsuji Nakamura, Toru Nagasawa, Fujio Yu, Ichiro Watanabe and Hideaki Yamada.: "A new enzymatic synthesis of (R)-gamma-chloro-beta-hydroxybutyronitrile." Tetrahedron.50. 11821-11826 (1994)

Tetsuji Nakamura、Toru Nagasawa、Fujio Yu、Ichiro Watanabe 和 Hideaki Yamada：“(R)-γ-氯-β-羟基丁腈的新酶法合成。”

Toru Nagasawa and Hideaki Yamada.: "Microbial production of commodity chemicals" Pure & Applied Chem.(in press). (1995)

Toru Nagasawa 和 Hideaki Yamada.：“日用化学品的微生物生产” Pure

Tetsuji Nakamura: "Characterization of a novel enantioselective halohydrin hydrogen-halide-lyase." Appl.Environ.Microbiol.60(in press). (1994)

Tetsuji Nakamura：“一种新型对映选择性卤代醇卤化氢裂解酶的表征。”

Tetsuji Nakamura: "Purification and characterization of two epoxide hydrolases from Corynebacterium sp.strain N-1074" J.Bacteriol.176(in press). (1994)

Tetsuji Nakamura：“来自棒状杆菌属菌株 N-1074 的两种环氧化物水解酶的纯化和表征”J.Bacteriol.176（出版中）。

TORU NAGASAWA: "Purification and Characterization of halohyduin hydrogen-halide-lyase from a recombiuant Escherichis Celi having its gene frew a Couyvehacteviums" Appl.Microbiol.Biotechnol.36. 478-482 (1992)

TORU NAGASAWA：“来自具有 Couyvehacteviums 基因的重组大肠杆菌的卤化氢卤化酶的纯化和表征”Appl.Microbiol.Biotechnol.36。