产β-淀粉酶菌株的筛选及β-淀粉酶基因在大肠杆菌中的克隆与表达  

The Isolation of Beta-amylase-producing Bacteria and Cloning,Expression of Beta-amylase Gene in Escherichia coli

作　　者：李猛[1] 陈利飞[1] 杨建楼 马春玲[1] 

机构地区：[1]齐鲁工业大学食品与生物工程学院山东省微生物工程重点实验室,济南250000

出　　处：《生物技术通报》2014年第12期161-167,共7页Biotechnology Bulletin

基　　金：山东省中青年科学家科研奖励基金项目(C010302)

摘　　要：从淀粉厂附近的土壤中筛选到一株能够利用淀粉的细菌,对该细菌进行生理生化检测和16S r DNA同源性比对,分析该菌株为蜡状芽孢杆菌(Bacillus cereus)。利用基因克隆技术得到了该菌株的β-淀粉酶基因,该基因含有一个约30个氨基酸的信号肽序列。将该β-淀粉酶基因重组进入质粒p ET-28a中,转化进入E.coli BL21(DE3)中进行表达。检测表达结果显示得到了重组后的β-淀粉酶蛋白质,重组酶的酶活力提高了53.9%。A bacterium which can degrade starch was isolate from the soil near the starch factory, was identified as Bacillus cereus by means of morphology and physiological-chemical characterization as well as 16S rDNA sequencing. Beta-amylase gene was cloned from the strain by PCR, and gene sequence was analyzed, which can encode a signal peptide which contains about 30 amino acids. The beta-amylase gene was cloned and expressed in Escherichia coli BL21 （ DE3 ） . The results showed that recombinant bacteria can produce beta-amylase, and enzymatic activity increased by 53.9% compared with the parent.

关 键 词：Β-淀粉酶 蜡状革孢杆菌 克隆 表达 

分 类 号：Q78[生物学—分子生物学]