热带海洋学报 ›› 2018, Vol. 37 ›› Issue (4): 38-44.doi: 10.11978/2017106

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深海来源微生物乙酰酯酶的酶学性质鉴定及拆分制备D-乳酸甲酯

黄锦龙1,3(), 张继福2(), 胡洁莹4, 关见留4, 张云1,3, 孙爱君1,3, 胡云峰1,3()   

  1. 1. 中国科学院南海海洋研究所, 中国科学院热带海洋生物资源与生态重点实验室, 广东 广州510301
    2. 广东省中医院, 广东 广州 510120
    3. 中国科学院南海海洋研究所, 广东省海洋药物重点实验室, 广东 广州 510301
    4. 广东第二师范学院, 广东 广州 510303
  • 收稿日期:2017-09-29 修回日期:2017-10-25 出版日期:2018-07-20 发布日期:2018-07-16
  • 作者简介:

    作者简介:黄锦龙(1986—), 男, 博士研究生, 研究方向为生物催化与酶工程。E-mail: jl_huang@163.com

  • 基金资助:
    中国科学院战略性先导科技专项项目(XDA11030404);广东省海洋渔业科技攻关与研发方向项目(A201701C12);广州市科技计划项目(201510010012)

Characterization of one deep-sea derived microbial acetyl esterase and its utilization in the preparation of D-methyl lactate through kinetic resolution

Jinlong HUANG1,3(), Jifu ZHANG2(), Jieying HU4, Jianliu GUAN4, Yun ZHANG1,3, Aijun SUN1,3, Yunfeng HU1,3()   

  1. 1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
    2. Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China
    3. Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
    4. Guangdong University of Education, Guangzhou 510303, China
  • Received:2017-09-29 Revised:2017-10-25 Online:2018-07-20 Published:2018-07-16
  • Supported by:
    Strategic Priority Research Program of the Chinese Academy of Sciences (XDA11030404);Scientific and Technological Project of Ocean and Fishery from Guangdong Province (A201701C12);Guangzhou Science and Technology Plan Projects (201510010012)

摘要:

D-乳酸及其酯是重要的手性药物中间体和手性化工产品。从南海深海芽孢杆菌Bacillus sp. SCSIO15029克隆到一个乙酰酯酶基因bae02030, 表达并鉴定该酶Bae02030的酶学性质。该酯酶的最适pH和最适温度分别为8.5和35℃, 其对多种有机溶剂和表面活性剂具有较好的耐受性。乙酰酯酶Bae02030能够通过水解拆分消旋乳酸甲酯来制备光学纯的D-乳酸甲酯。通过对拆分反应进行优化, 添加体积分数为60%的正庚烷能够改善乙酰酯酶Bae02030的光学选择性, 所制备的D-乳酸甲酯的对映体过量值(e.e.s)超过99%, 转化率(c)为56%。深海微生物来源的乙酰酯酶Bae02030作为生物催化剂在工业上制备手性药物中间体具有较好的应用潜力。

关键词: 深海微生物, 乙酰酯酶, 生物催化, D-乳酸甲酯, 手性拆分

Abstract:

D-lactic acid and its esters are important chiral drug intermediates and chemicals. An acetyl esterase, Bae02030, from Bacillus sp. SCSIO15029 isolated from the deep sea of the South China Sea was cloned, expressed and functionally characterized. The optimum pH and temperature of acetyl esterase Bae02030 were 8.5 and 35°C, respectively. Bae02030 exhibited excellent resistance to most organic solvents and surfactants tested. Bae02030 could generate optically pure D-methyl lactate through resolution of racemic methyl lactate. After process optimization of the enzymatic resolution reactions, the addition of 60% heptane improved the enantio-selectivity of Bae02030. The enantiomeric excess of prepared D-methyl lactate was over 99% and conversion rate reached 56%. Acetyl esterase Bae02030 identified from deep-sea microorganism, as a biocatalyst, possesses great potential in the production of chiral drug intermediates in industry.

Key words: deep-sea microorganisms, acetyl esterase, biocatalysis, D-methyl lactate, chiral resolution

中图分类号: 

  • P735.51