Functional Analyses and Application of Microbial Lactonohydrolases

微生物内酯水解酶的功能分析及应用

• 批准号: 14360054
• 负责人: KATAOKA Michihiko
• 金额: $ 9.6万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360054/
• 关键词: lactonase; lactone; Fusarium oxysporum; Agrobacterium tumefaciens; Optical resolution; Lactone-ring cleaving enzyme; Acinetobacter calcoaceticus; Asymmetric hydrolysis

Microbial lactonohydrolases (intramolecular ester bond-hydrolyzing enzymes) with unique properties were found and characterized. The lactonohydrolase from Fusarium oxysporum catalyzes enantioselective hydrolysis of aldonate lactones and D-pantoyl lactone (D-PL). This enzyme is useful for the large-scale optical resolution of racemic PL.. The gene encoding latonohydrolase of F. oxysporum could be overexpressed in Aspergillus oryzae.The Agrobacterium tumefaciens enzyme catalyzes, asymmetric hydrolysis of PL, but the stereospecificity is opposite to that of the Fusarium enzyme. This enzyme gene also, could be expressed in E. coli transformant cells.Dihydrocoumarin hydrolase (DCH) from Acinetobacter calcoaceticus is a bifunctional enzyme, which catalyzes not only hydrolysis of aromatic lactones but also bromination of monochlorodirnedon in the presence of H_2O_2 and dihydrocoumarin. DCH also hydrolyzes several linear esters, and is useful for enantioselective hydrolysis of methyl DL-β-acetylthioisobutyrate and regioselective hydrolysis of methyl cetraxate. DCH in vivo detoxified peroxoacids in conjunction with the catalase.

发现并鉴定了具有独特性质的微生物内酯水解酶（分子内酯键水解酶）。尖孢镰刀菌（Fusarium oxysporum）的内酯水解酶催化醛糖酸内酯和D-泛酰内酯（D-PL）的对映选择性水解。该酶可用于大规模光学拆分外消旋PL。对F.根癌农杆菌酶催化PL的不对称水解，但立体专一性与镰刀菌酶相反。该酶基因也可在大肠杆菌中表达。乙酸钙不动杆菌（Acinetobacter calcoaceticus）的二氢香豆素水解酶（DCH）是一种双功能酶，在H_2O_2和二氢香豆素存在下，它不仅能催化芳香内酯的水解，还能催化一氯二烯酮的溴代反应。DCH还水解几种直链酯，并且可用于DL-β-乙酰基硫代异丁酸甲酯的对映选择性水解和西曲酸甲酯的区域选择性水解。DCH在体内解毒过氧酸与过氧化氢酶。

项目成果

K.Honda et al.: "A novel tool for chiral-technology "Lactonase" (in Japanese)"Bioscience & Industry. 60. 163-166 (2002)

K.Honda 等人：“一种新型手性技术工具“内酯酶”（日语）”生物科学

K.Honda et al.: "Biocatalytic deprotection of a cetraxate ester by Microbacterium sp. 7-1W cells"Biosci.Biotechnol.Biochem.. 66. 192-194 (2003)

K.Honda 等人：“微杆菌属 sp. 7-1W 细胞对西曲酯的生物催化脱保护”Biosci.Biotechnol.Biochem.. 66. 192-194 (2003)

K.Honda et al.: "Biocatalytic department of a cetraxate ester by Microbacterium sp. strain 7-1W cells"Biosci.Biotechnol.Biochem.. 67・1. 192-194 (2003)

K. Honda 等：“微杆菌菌株 7-1W 细胞对西曲酯的生物催化作用”Biosci.Biotechnol.Biochem.. 67・1 (2003)。

K.Honda et al.: "Role of Acinetobacter calcoaceticus 3,4-dihydrocoumarin hydrolase in oxidative stress defence against peroxoacids"Eur.J.Biochem.. 270. 486-494 (2003)

K.Honda 等：“乙酸钙不动杆菌 3,4-二氢香豆素水解酶在针对过氧酸的氧化应激防御中的作用”Eur.J.Biochem.. 270. 486-494 (2003)

K.Honda et al.: "Enzymatic preparation of D-b-acetylisobutyric acid and cetraxate hydrochloride using a stereo-and/orregioselective hydrolase, 3, 4-dihydrocoumarin hydrolase, from Acinetobacter calcoaceticus"Appl.Microbiol.Biotechnol.. 60. 288-292 (2002)

K.Honda 等人：“使用来自醋酸钙不动杆菌的立体和/或区域选择性水解酶、3, 4-二氢香豆素水解酶，酶法制备 D-b-乙酰基异丁酸和盐酸西曲沙酯”Appl.Microbiol.Biotechnol.. 60. 288-292