多粘类芽孢杆菌β-葡萄糖苷酶bglA、bglB和rbgl基因
在大肠杆菌中的表达
王艳;马亚茹;万学瑞;王川;吴润;刘桂林;刘原子;吴自祥
【期刊名称】《草业学报》 【年(卷),期】2017(026)005
【摘要】To improve the enzyme activity of β-glucosidase,twoβ-glucosidase genes from Bacilluspolymyxa were introduced separately (bglAandbglB)and together (bgl)into the pET-28a vector and expressed in Esch-erichiacoliC41 .The three recombinant strains were designated as EA,EB,and co-expression EAB.Strains EA and EB were mixed at ratios of 1:1 ,1:2 ,and 2:1 and their totalβ-glucosidase activity was compared with those of each single enzyme group,the co-expression strain,and mixed expression strains.The results of SDS-PAGE analyses showed that both BglA and BglB were 50 ku,and the size of these proteins in the EA and EB mixed cultures was also 50 ku.The size of Bgl in the co-expression strain EAB was 100 ku.These results indicated that a Bgl complex was able to form in cells,but notinvitro.In an enzyme activity assay,the activi-ty of Bgl from the co-expression strain was not significantly different from that ofbglin B.polymyxa,but it was significantly higher than those of BglA in EA and BglB in EB (P<0.05).The results of Congo red stai-ning also showed that the enzyme activity of Bgl was significantly higher than those of BglA and BglB.This study lays
多粘类芽孢杆菌β-葡萄糖苷酶bglA、bglB和rbgl基因在大肠杆菌中的表达
