麦芽糖诱导软化芽孢杆菌α-环糊精葡萄糖基转移酶在枯草杆菌中的表达  

Expression of Paenibacillus macerans α-cyclodextrin Glycosyltraferase in Bacillus subtilis by Maltose Induction

作　　者：张佳瑜[1,2] 吴丹[1,2] 陈晟[1,2] 陈坚[1,2] 吴敬[1,2] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,无锡214122 [2]江南大学生物工程学院工业生物技术教育部重点实验室,无锡214122

出　　处：《中国生物工程杂志》2010年第12期42-48,共7页China Biotechnology

基　　金：国家杰出青年基金(20625619);科技部食品科学与技术国家重点实验室科研基金(SKLF-MB-200802);中央高校基本科研业务费专项资金(JUSRP20917)资助项目

摘　　要：通过PCR扩增软化芽孢杆菌α-环糊精葡萄糖基转移酶基因,将基因片段克隆到大肠杆菌-枯草杆菌穿梭载体pGJ103中,转化枯草杆菌WB600得基因工程菌进行外源表达。在1.5%的麦芽糖初始发酵培养基上摇瓶培养,48 h后重组枯草杆菌产酶活性为6.1U/ml。通过单因素分析和响应面分析对重组枯草杆菌产CGT酶摇瓶发酵条件进行优化。分析得到培养基关键组分麦芽糖,玉米淀粉和酵母粉三者最佳浓度分别为:15.5g/L,13g/L和20g/L。在此条件下,摇瓶培养36h后α-CGT酶活性为17.6U/m l,5L罐分批发酵30h后酶活达到20U/ml(水解活性为1.4×104IU/ml)。The α-cgt gene was obtained from Paenibacillus macerans by PCR,and then it was cloned into Escherichia coli-Bacillus subtilis shuttle vector pGJ103 and transformed into B.subtilis WB600.After cultivation for 48 h with shake flask in 1.5% maltose initial medium,the α-CGTase activity of recombinant B.subtilis was 6.1U/ml.In addition,the experiment optimized the culture conditions of the recombinant B.subtilis strain in shaking flask by means of a single factor synthesis and Box-Behnken design（BBD）.The analysis predicted the concentrations of maltose,corn starch and yeast extract were at 15 g/L,13 g/L,and 20 g/L,respectively.In this condition,the experimental result of 17.6 U/ml could be obtained when the cells were cultured for 36 h in shaking flasks.The CGTase activity reached 20 U/ml at 30 h of culture in a 5 L bioreactor（hydrolysis activity was 1.4×104 IU/ml）.

关 键 词：α-CGT酶 枯草杆菌 外源表达 响应面优化 发酵 

分 类 号：Q786[生物学—分子生物学]