Aspergillus ficuum内切菊粉酶基因在大肠杆菌中表达  

Expression of Endo-Inulinase gene from Aspergillus ficuum in Escherichia coli

作　　者：陈晓明[1,2] 陈静 陈寒青 金征宇 徐学明 

机构地区：[1]食品科举与技术国家重点实验宦,江南大学,江苏无锡214122 [2]江南大学食品学院,江苏无锡214122

出　　处：《食品与生物技术学报》2011年第3期388-393,共6页Journal of Food Science and Biotechnology

基　　金：食品科学与技术国家重点实验室2008年度目标导向项目(SKLF-MB-200804);江苏省自然科学基金创新学者攀登项目(BK2008003)

摘　　要：研究内切菊粉酶基因在大肠杆菌中表达,利用基因工程的原理和方法,将来源于Asper-gillus ficuum JNSP5-06的内切菊粉酶基因(endoI)克隆到pET-28a(+),并在大肠杆菌BL21(DE3)中进行表达。重组菌经IPTG诱导后对表达产物进行酶活检测和酶水解产物分析。结果表明已成功构建表达载体pET28a-endoI,表达后的粗酶活为35 U/mL发酵液;重组酶水解菊粉的产物经TLC分析证实酶液具有内切菊粉酶活性,水解产物主要为低聚二糖、三糖和四糖。为其应用和工业化生产奠定基础。In order to expand the industrial application of endo-inulinase,its gene from Aspergillus ficuum JNSP5-06 was cloned into the pET-28a（＋） and expressed in E.coli BL21（DE3） and get a mutant strain.The mutant was induced by IPTG before the hydrolysis activity was measured and the product was analyzed.The expression vector pET28a-endoI was successfully obtained and the crude enzyme activity was 35 U/mL（fermentation broth）.The hydrolysis product from the recombinant inulinase was identified by TLC analysis,indicating that endo-inulinase enzyme solution had activity and the major products were disaccharide,trisaccharide,and tetrasaccharide.These results suggested that the endo-inulinase gene has been successfully expressed in E.coli and significantly improved the endo-inulinase activity.

关 键 词：内切菊粉酶基因 克隆 表达 低聚果糖 

分 类 号：Q78[生物学—分子生物学]