环氧树脂固定化的Bacillus sp.DL-2胞外蛋白酶在拆分(±)-乙酸苏合香酯中的应用  

Immobilization of Extracellaluar Proteases of Bacillus sp.DL-2 Using Epoxy Resin to Asymmetrically Hydrolyze(±)-1-Phenylethyl Acetate

作　　者：董璐 张继福[4] 张云[1] 胡云峰[1,3] DONG Lu;ZHANG Ji-fu;ZHANG Yun;HU Yun-feng(Key Laboratory of Tropical Marine Bio-resources and Ecology,Guangdong Key Laboratory of Marine Materia Medical,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China;University of Chinese Academy of Sciences,Beijing 100049,China;Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou),Guangzhou 511458,China;Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510120,China)

机构地区：[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省海洋药物重点实验室,广州510301 [2]中国科学院大学,北京100049 [3]南方海洋科学与工程广东省实验室(广州),广州511458 [4]广东省中医院,广州510120

出　　处：《中国生物工程杂志》2020年第4期49-58,共10页China Biotechnology

基　　金：南方海洋科学与工程广东省实验室(广州)人才团队引进重大专项(GML2019ZD0406);广东省自然科学基金项目(2018A030313151);广东省海洋渔业科技攻关与研发方向项目(A201701C12);中国科学院战略性先导科技专项(XDA11030404);中国科学院“科学”号高端用户项目(KEXUE2018G05)资助项目.

摘　　要：环氧基是一个非常活跃的基团,它能与酶、蛋白质和核酸等生物分子发生反应形成共价键,有利于生物分子的固定化。经共价结合法固定化的酶其稳定性及重复使用性可得到显著提高。用环氧树脂ES-103B为载体采用共价结合法对海洋细菌Bacillus sp.DL-2的胞外蛋白酶进行固定化,经过单因素实验优化条件得出最优固定化条件为:p H 8.0的胞外蛋白酶溶液,25 g/L的ES-103B,45℃下反应8h。采用此最优条件下的固定化酶拆分(±)-乙酸苏合香酯制备出了e.e.p=97.5%的(R)-1-苯乙醇(产率为45.0%)和e.e.s=99.2%的(S)-乙酸苏合香酯(产率为83.9%)。该固定化酶拆分(±)-乙酸苏合香酯在重复使用8次后制备出的(R)-1-苯乙醇的e.e.p仍大于90%,且固定化胞外蛋白酶在4℃下具有较好的储存稳定性。Epoxy group,a very active group,can react with biomolecules such as enzymes,proteins,nucleic acid and form covalent bonds,which is beneficial for the immobilization of biomolecules.Enzymes immobilized on carriers via covalent binding exhibit better stability and reusability than free enzymes.The extracellular proteases of marine bacterium Bacillus sp.DL-2 were immobilized by covalent bonding onto epoxy resin ES-103 B.After the the optimization of immobilization conditions,pH 8.0 extracellular proteases solution,25 g/L epoxy resin ES-103 B,and 45℃for 8 h were determined as the optimal conditions for immobilized proteases to asymmetrically hydrolyze(±)-1-phenylethyl acetate,which generated(R)-1-phenylethanol with the e.e.p being 97.5%and the yield being 45.0%,and generated(S)-1-phenylethyl acetate with the e.e.s being99.2%and the yield being 83.9%,respectively.The the immobilized enzyme could be reused to resolve(±)-1-phenylethyl acetate for eight times and the e.e.p still retained over 90%.Additionally,the immobilized extracellular proteases showed good storage stability at 4℃.

关 键 词：环氧树脂ES-103B 海洋微生物 固定化酶 不对称水解 (R)-1-苯乙醇 (S)-乙酸苏合香酯 

分 类 号：Q502[生物学—生物化学]