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黑曲霉(Aspergillus niger)ZJB-09101菌株手性拆分环氧氯丙烷的培养条件优化
引用本文:陈月园,许辉辉,胡忠策,郑裕国.黑曲霉(Aspergillus niger)ZJB-09101菌株手性拆分环氧氯丙烷的培养条件优化[J].浙江化工,2009,40(8):11-16.
作者姓名:陈月园  许辉辉  胡忠策  郑裕国
作者单位:浙江工业大学,生物工程研究所,浙江,杭州,310014
摘    要:将筛选得到的黑曲霉(Aspergillus niger)ZJB-09101菌株应用于外消旋环氧氯丙烷的手性拆分,对菌种产环氧化物水解酶的发酵条件和外消旋环氧氯丙烷的手性催化过程进行了研究。结果表明,黑曲霉ZJB-09101产环氧化物水解酶的最佳培养基成分是:淀粉14.63g/L,豆饼粉6.45g/L,K2HPO4·3H2O0.8g/L,KH2PO4·3H2O0.4g/L,MaSO4·7H2O0.05g/L,pH7.0。ZJB-09101菌株在150r/min、30℃条件下,经过4d的培养,环氧化物水解酶的酶活达90.36U/1,,比优化前的39.27U/L提高了130%。在环己烷的反应体系中,底物外消旋环氧氯丙烷浓度为0.6%,在30℃下生物催化反应7h,(S)-环氧氯丙烷的e.e.值和产率分别为99.060/6:和17.14%。

关 键 词:手性环氧氯丙烷  黑曲霉  环氧化物水解酶  响应面法

Optimization of Cultivation Conditions of Aspergillus Niger ZJB-09101 for Enzymatic Resolution of Racemic Epichlorohydrin
CHEN Yue-Yuan,XU Hui-Hui,HU Zhong-Ce,ZHENG Yu-Guo.Optimization of Cultivation Conditions of Aspergillus Niger ZJB-09101 for Enzymatic Resolution of Racemic Epichlorohydrin[J].Zhejiang Chemical Industry,2009,40(8):11-16.
Authors:CHEN Yue-Yuan  XU Hui-Hui  HU Zhong-Ce  ZHENG Yu-Guo
Affiliation:(Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China)
Abstract:The isolated strain, Aspergillus niger ZJB-090101, was employed in this work for enzyme resolution of racemie epichlorohydrin. The cultivation conditions of A. niger ZJB-09101 for production of epoxide hydrolase, and the enzymatic characteristics of epoxide hydrolase were investigated. The results showed that the optimal medium for enzyme production was as follows: starch 14.63 g/L, soybean cake 6.45 g/L, KH2PO4 0.4 g/L, K2HPO4 0.8 g/L, MgSO4 .7H20 0.05 g/L, and initial culture pH was 7.0. The maximal enzyme activity of 90.36 U/L was obtained after 4 days cultivation at 30℃ with 150 r/min, and increased by 130% as compared with that of unoptimized culture condition. The enzymatic resolution was carried out at cyclohexane solution with 0.6% of racemic epichlorohydrin at 30℃. After 7 hours reaction, entanselect excess (e.e.) and product yield of (S)- epichlorohydrin reached 99.06% and 17.14%, respectively.
Keywords:chiral epichlorohydrin  aspergillus niger ZJB-090101  epoxide hydrolase  response surface methodology (RSM)
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