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通过展示毕赤酵母全细胞生物催化剂的南极假丝酵母脂肪酶 B 催化的 sec-醇的动力学拆分。

Kinetic resolution of sec-alcohols catalysed by Candida antarctica lipase B displaying Pichia pastoris whole-cell biocatalyst.

机构信息

Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, PR China.

Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, PR China.

出版信息

Enzyme Microb Technol. 2018 Mar;110:8-13. doi: 10.1016/j.enzmictec.2017.11.005. Epub 2017 Nov 21.

Abstract

Kinetic resolution of sec-alcohols is a green process with biocatalyst. Candida antarctica lipase B (CALB) displayed on Pichia pastoris cell-surface (Pp-CALB) was characterized in kinetic resolution of sec-alcohols with different structures. The reaction parameters including acyl donors, molar ratio of substrates, solvents and temperatures were examined with 2-octanol as model substrate. 47.4% molar conversion of 2-octanol and 99.7% ee were obtained after a 5h reaction with Pp-CALB, and 90% of its original activity still remained after being reused for 10 cycles. Pp-CALB was then used to several sec-alcohols and it showed great enzymatic activity and enantioselectivity to all tested sec-alcohols, more than 93.1% of ee. The enantioselective characteristics of Pp-CALB catalysed sec-alcohols with different structures were compared with Novozyme 435 which was almost the same. Solvent free system as one way of green chemistry was applied to Pp-CALB and Pp-CALB showed great catalytic activity and enantioselectivity. Pp-CALB was potential biocatalyst of high enzymatic activity and enantioselectivity using in resolution of sec-alcohols.

摘要

手性醇的动力学拆分是一种绿色过程,使用生物催化剂。在不同结构的仲醇动力学拆分中,展示在毕赤酵母细胞表面的南极假丝酵母脂肪酶 B(CALB)(Pp-CALB)的特性。以 2-辛醇为模型底物,考察了酰基供体、底物摩尔比、溶剂和温度等反应参数。用 Pp-CALB 反应 5 小时后,2-辛醇的摩尔转化率为 47.4%,ee 值为 99.7%,重复使用 10 次后,其原始活性仍保留 90%。然后,将 Pp-CALB 用于几种仲醇,它对所有测试的仲醇都表现出很高的酶活性和对映选择性,ee 值均超过 93.1%。比较了具有不同结构的仲醇在手性醇的对映选择性特征,与 Novozyme 435 几乎相同。无溶剂体系作为绿色化学的一种方法被应用于 Pp-CALB,Pp-CALB 表现出很高的催化活性和对映选择性。Pp-CALB 是一种潜在的高酶活性和对映选择性的生物催化剂,可用于仲醇的拆分。

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