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新型微生物脂肪酶:在有机介质反应中的催化活性

Novel microbial lipases: catalytic activity in reactions in organic media.

作者信息

Cardenas F, de Castro M S., Sanchez-Montero J M., Sinisterra J V., Valmaseda M, Elson S W., Alvarez E

机构信息

SmithKline Beecham, Centro de Investigación Básica, Santiago Grisolía, 4, Parque Tecnológico de Madrid, 28760 Tres Cantos, Madrid, Spain

出版信息

Enzyme Microb Technol. 2001 Feb 1;28(2-3):145-154. doi: 10.1016/s0141-0229(00)00278-7.

Abstract

2,000 microbial strains were isolated from soil samples and tested to determine their lipolytic activity by employing screening techniques on solid and in liquid media. Culture broths were initially tested with 1,2-O-dilauryl-rac-glycero-3-glutaric acid-resorufinyl ester, a chromogenic substrate specific for lipases. Fourteen lipase-producing microorganisms were selected and their taxonomic identification was carried out. Hydrolysis of tributyrin or olive oil and the esterification of oleic acid with heptanol were selected to preliminary evaluate the catalytic activity of these lipases. All the selected lipases catalysed this esterification reaction with good yields. Resolution of (R,S)-2-(4-isobutylphenyl) propionic acid, (R,S)-1-phenylethanol, (R,S) 1-phenylethylamine and of (R) or (S) glycidol were performed to evaluate the stereoselectivity of these novel enzymes as biocatalysts in reactions in organic media. Lipases from the fungi Fusarium oxysporum and Ovadendron sulphureo-ochraceum gave the best yields and enantioselectivities in the resolution of racemic ibuprofen and 1-phenylethanol. Several lipases displayed a high stereoselectivity in the resolution of chiral amines by an alcoxycarbonylation reaction.

摘要

从土壤样品中分离出2000株微生物菌株,并通过在固体和液体培养基上采用筛选技术来测试它们的脂肪分解活性。培养肉汤最初用1,2-O-二月桂酰-外消旋甘油-3-戊二酸-试卤灵酯进行测试,这是一种对脂肪酶具有特异性的显色底物。选择了14株产脂肪酶的微生物并进行了分类鉴定。选择三丁酸甘油酯或橄榄油的水解以及油酸与庚醇的酯化反应来初步评估这些脂肪酶的催化活性。所有选定的脂肪酶都能以良好的产率催化这种酯化反应。进行了(R,S)-2-(4-异丁基苯基)丙酸、(R,S)-1-苯乙醇、(R,S)-1-苯乙胺以及(R)或(S)缩水甘油的拆分,以评估这些新型酶作为有机介质中生物催化剂的立体选择性。来自尖孢镰刀菌和硫黄赭色奥瓦德菌的脂肪酶在消旋布洛芬和1-苯乙醇的拆分中给出了最佳产率和对映选择性。几种脂肪酶在通过烷氧基羰基化反应拆分手性胺时表现出高立体选择性。

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