β-葡萄糖苷酶高产菌株的筛选及其基因的克隆与表达  

Screening of A Strain for High β-Glucosidase Producing and Cloning, Expression of Its β-Glucosidase Gene

作　　者：王斌斌 刘逸寒[1] 李玉[1] 王建玲[1] 路福平[1] 

机构地区：[1]工业发酵微生物教育部重点实验室(天津科技大学)工业酶国家工程实验室天津市工业微生物重点实验室天津科技大学生物工程学院,天津300457

出　　处：《化学与生物工程》2012年第6期66-70,共5页Chemistry & Bioengineering

摘　　要：利用栀子苷培养基从滨海新区盐碱地土样中筛选得到一株高产β-葡萄糖苷酶菌株,酶活力达到14.82U·mL-1,经16SrDNA鉴定,命名为短小芽孢杆菌B-4。克隆获得B-4β-葡萄糖苷酶基因,测序结果表明,其大小为1437bp,与GenBank中短小芽孢杆菌SAFR-032β-葡萄糖苷酶基因YP_001488769.1序列比对,核苷酸序列同源性达97%,氨基酸序列同源性达99%。进一步利用表达载体pET-22b(+),实现β-葡萄糖苷酶基因bglB在大肠杆菌BL21(DE3)中的高效表达,酶活力达46.85U·mL-1。A strain which could produce β-glucosidase was screened by jasminoidin medium from soil samples and it possessed high β-glueosidase activity（14.82 U . mL^-1）. It was named as Bacillus pumilus B-4 after de- termination of 16S rDNA. Theβ-glucosidase gene was cloned from Bacillus pumilus SAFR-032 gene（bglB） YP _001488769. 1 with PCR method. The sequencing results showed that the cloned gene fragment was 1437 bp. Compared the sequence with that of glucosidase gene YP_001488769. 1, the homology of nucleotide sequence and amino acid sequence reached 97 %, 99%, respectively. Then the expression vector pET-bgIB was constructed and transformed into E. coli BL21（DE3）. Finally,it was successfully expressed in E. coli and the maximum glucosidase activity（46.85 U . mL^-1） was achieved.

关 键 词：短小芽孢杆菌 Β-葡萄糖苷酶 克隆 大肠杆菌 表达 

分 类 号：Q78[生物学—分子生物学] Q556.2