中国农业大学学报2012，Vol.17Issue(5)：92-98,7.
纤维素酶高产菌的筛选鉴定及纤维素酶基因的克隆与表达
Cloning and expression of a novel cellulase gene from an solated Bacillus pumilus strain
摘要
Abstract
The aim of this study was to exploit new resources of cellulolytic bacteria and new cellulose genes for industrial processes.In this study,targeted bacteria were screened by CMC amended BHM medium and identified by Gram staining with 16S rDNA partial sequence alignment.After amplification of cellulase gene using primer pair P,it was ligated to pET-28a(+) in corresponding sites to construct prokaryotic expression vector.Heterogeneous expression was conducted in E.coli BL21(DE3) and verified by SDS-PAGE and enzyme detection.The Bacillus pumilus BY-1 with premium enzyme activity was successfully screened.Under the optimum condition of 55 ℃ in temperature and 7.45 in pH,the crude enzyme,reached the maximum enzyme activity at 0.921 6 μmol/(mL·min).Simultaneously,a new 1 851 bp cellulase gene bglC-BY with the structure of GH9/CBM3 was successfully cloned from this bacterial strain.After IPTG induction,SDS-PAGE verified the successful expression of bglC-BY with a mass of about 70 u and the enzyme activity was 0.541 3 μmol/(mL·min).Thus,a potential new strain and a new endoglucanase gene with premium activity can be provided for industrial processes.关键词
纤维分解菌/筛选/纤维素酶基因/克隆/表达Key words
cellulolytic microbes/screening/cellulase gene/cloning/expression分类
农业科技引用本文复制引用
刘海艳,白龙,赵海波,杨明明,曹斌云..纤维素酶高产菌的筛选鉴定及纤维素酶基因的克隆与表达[J].中国农业大学学报,2012,17(5):92-98,7.基金项目
国家科技支撑计划项目 （）
陕西省科技统筹创新工程计划 （）
