| 木薯醇腈酶在枯草芽孢杆菌中的表达及活性检测 |
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| 引用本文： | 张冬冬,郭晓军,刘慧娟,雷白时,牛振东,朱宝成.木薯醇腈酶在枯草芽孢杆菌中的表达及活性检测[J].河北农业大学学报,2009,32(3). |
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| 作者姓名： | 张冬冬 郭晓军 刘慧娟 雷白时 牛振东 朱宝成 |
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| 作者单位： | 1. 河北农业大学生命科学学院,河北保定,071001
2. 中国科学院微生物研究所,北京,100091 |
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| 摘 要： | 醇腈酶(α-hydroxynitrile lyase,HNL)能够催化羰基化合物和HCN立体选择性的加工形成手性醇腈化合物。以含有木薯醇腈酶基因的重组质粒pET28a-HNL为模板,应用PCR扩增得到HNL基因,将其连接到pMD18-T载体中进行测序分析,然后通过Nde I和Xba I2个酶切位点将其连接到枯草芽孢杆菌表达载体pMA5Z2中,获得了含有HNL基因的重组质粒pMA5Z2-HNL,转化蛋白质三缺陷的枯草芽孢杆菌DB1342。SDS-PAGE显示HNL在枯草芽孢杆菌DB1342中获得表达,产物分泌到细胞外,每毫升发酵液中获得了2.108 U的醇腈酶。
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| 关 键 词： | 木薯醇腈酶 枯草芽孢杆菌 分泌表达 活性 |
Expression and characterization of hydroxynitrile lyase from Manihot esculenta in Bacillus subtilis |
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| ZHANG Dong-dong,GUO Xiao-jun,LIU Hui-juan,LEI Bai-shi,NIU Zhen-dong,ZHU Bao-cheng.Expression and characterization of hydroxynitrile lyase from Manihot esculenta in Bacillus subtilis[J].Journal of Agricultural University of Hebei,2009,32(3). |
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| Authors: | ZHANG Dong-dong GUO Xiao-jun LIU Hui-juan LEI Bai-shi NIU Zhen-dong ZHU Bao-cheng |
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| Institution: | 1.College of life Science;Agricultural University of Hebei;Baoding 071001;China;2.Institute of Microbiology;Chinese Academy of Sciences;Beijing 100091;China |
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| Abstract: | Hydroxynitrile lyase(HNL) can catalyze carbonyls and HCN which stereoselectively manufacture and form chiral cyanohydrin components.HNL gene was amplified by PCR using plasmid pET28a-HNL which contain the gene of hydroxynitrile lyase from Manihot esculenta as template and then ligated into pMD18-T vector in order to execute sequence analysis.Then the amplified DNA fragment was ligated into the Bacillus subtilis expression vector pMA5Z2 using Nde I and Xba I.The recombinated plasmid pMA5Z2-HNL was obtained a... |
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| Keywords: | hydroxynitrile lyase from Manihot esculenta Bacillus subtilis secreted expression activity |
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