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 produceβ-glucosidase was screened by jasminoidin medium from soil samples and it possessed highβ-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.
作者
王斌斌
刘逸寒
李玉
王建玲
路福平
WANG Bin-bin;LIU Yi-han;LI Yu;WANG Jian-ling;LU Fu-ping(Laboratory of Industrial Fermentation Microbiology,Tianjin University of Science&Technology,Ministry of Education,National Engineering Laboratory for Industrial Enzymes,Tianjin Key Laboratory of Industrial Microbiology,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)
出处
《化学与生物工程》
CAS
2012年第6期66-70,共5页
Chemistry & Bioengineering
