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谷氨酸棒杆菌海藻糖合成酶基因的克隆及功能鉴定
引用本文:韦宇拓,朱绮霞,罗兆飞,陈发忠,汪嵘,黄日波.谷氨酸棒杆菌海藻糖合成酶基因的克隆及功能鉴定[J].工业微生物,2005,35(2):1-6.
作者姓名:韦宇拓  朱绮霞  罗兆飞  陈发忠  汪嵘  黄日波
作者单位:1. 广西大学生命科学与技术学院,广西,南宁,530005
2. 南宁中诺生物工程公司,广西,南宁,530003
基金项目:国家高技术“863”资助项目(2001AA214171)
摘    要:利用生物信息学手段,在GenBank数据库进行氨基酸的同源性检索分析,发现来自谷氨酸棒杆茵(Corynebacterium glutamicum)一功能未确定的ORF序列被注释为假设的海藻糖酶(putative trehalose sesynthase),它与已报道的海藻糖合成酶的氨基酸序列有60%以上的同源性。本研究把这段ORF克隆到大肠杆茵进行表达及进行功能鉴定。实验表明这段ORF序列为一新的海藻糖合成酶基因,其表达产物能将麦芽糖分子转化成海藻糖分子。重组酶性质的初步研究表明重组酶在pH7.0~7.5,30℃转化麦芽糖效率最高。

关 键 词:谷氨酸棒杆菌  海藻糖合成酶  基因克隆  功能鉴定  海藻糖

Cloning and identification of novel gene encoding trehalose synthase from Corynebacterium glutamicum
WEI Yu-tuo,ZHU Qi-xia,LUO Zhao-fei,CHEN Fa-zhong,WANG Rong,HUANG Ri-bo.Cloning and identification of novel gene encoding trehalose synthase from Corynebacterium glutamicum[J].Industrial Microbiology,2005,35(2):1-6.
Authors:WEI Yu-tuo  ZHU Qi-xia  LUO Zhao-fei  CHEN Fa-zhong  WANG Rong  HUANG Ri-bo
Institution:WEI Yu-tuo~1,ZHU Qi-xia~2,LUO Zhao-fei~2,CHEN Fa-zhong,WANG Rong~1 HUANG Ri-bo~1
Abstract:An open reading frame (ORF) was annotated as putative trehalose synthase in Corynebacterium glutamicum sequenced genome, which has been found in GenBank database through amino homology blast base on combination of bioinformatics. Blasting against the database indicated that amino acid sequence of this ORF had less than 60% homology with the reported terhalose synthases. The function, which was identified by expressing this ORF in Escherichia coli, proved this ORF was a novel trehalose synthase gene. Recombinant enzyme showed the optimal activity was at pH 7.07.5 and 30 when it converted maltose into trehalose.
Keywords:Corynebacterium glutamicum  terhalose synthases  cloning  identified function
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