摘要
纳豆芽孢杆菌谷氨酸脱氢酶(GDH)是纳豆发酵产氨的关键酶。本文以纳豆芽孢杆菌基因组DNA为模板,根据枯草芽孢杆菌的GDH编码基因(RocG)序列设计引物,进行PCR扩增,再经pMD19-T质粒TA克隆,获得了全长为1275bp、与RocG基因同源性为98%的纳豆芽孢杆菌GDH的编码基因。构建原核表达载体pET28a-GDH,转化至E.coliBL21(DE3)进行体外诱导表达。表达的包涵体在超声破菌、变性、纯化和复性后,经SDS-PAGE分析得到了47kDa的特异条带。纳豆芽孢杆菌GDH同时具有NAD+和NADP+依赖活性,酶活分别为6.7723U·mg-1蛋白和9.4205U·mg-1蛋白。
Glutamate dehydrogenase of Bacillus Subtilis Natto is one of the key enzymes for ammonia production during natto fermentation.In order to clone and express the gene encoding the glutamate dehydrogenase of Bacillus Subtilis Natto,the PCR primers were designed on the published sequence of RocG gene of Bacillus Subtilis str168.The PCR product was cloned into vector pMD19-T and the result of gene sequencing exhibited a 98% similarity to RocG gene of Bacillus Subtilis str168.Thereafter,the gene was cloned into e...
出处
《上海交通大学学报（农业科学版）》
2010年第1期82-86,共5页
Journal of Shanghai Jiaotong University(Agricultural Science)
关键词
纳豆芽孢杆菌
谷氨酸脱氢酶
基因表达
酶活
Bacillus Subtilis Natto
glutamate dehydrogenase
gene expression
GDH activity
