农业环境科学学报Issue(4)：788-793,6.DOI:10.11654/jaes.2014.04.025
高温纤维素降解菌的筛选及内切酶基因的克隆表达
Isolation of Thermophilic Cellulose-Decomposing Bacteria and Cloning and Expression of an Endoglucanase Gene
摘要
Abstract
Thermophilic cellulose-decomposing bacteria could shorten digesting time of composts. Here we isolated eight strains of ther-mopilic cellulose-decomposing bacteria from composts produced from dairy manure and rice chaff using filter paper as a sole carbon source. After 48 hour cultivation on CMC-congo red medium, strains T1 and T2 had bigger clear zones than the other strains. Based on the morpho-logical, physiological and biochemical properties as well as 16S rRNA homology analysis, the strains T1 and T2 were identified as Bacillus cereus and Bacillus subtilis respectively. Endoglucanase gene of T2 was cloned and domains analysis showed that the endoglucanase gene contained an open reading frame of 1500 bp, corresponding to 499 amino acids. The endoglucanase was comprised of two discontinuous do-mains:one was N terminal catalytic domain belonging to glycosyl hydrolase family 5, and the other was C terminal substrate binding domain belonging to carbohydrate binding domain family 3. Using PET28a as the expression plasmid, the endoglucanase gene of strain T2 was suc-cessfully expressed in E. coli BL21(DE3). SDS-PAGE analysis showed that molecular weight of the expressed protein was about 50 KD and the concentration was about 93.14 mg·L-1.关键词
纤维素降解菌/内切酶基因/克隆/表达/结构域Key words
thermophilic bacteria/endoglucanase gene/cloning/expression/domain分类
资源环境引用本文复制引用
田伟,王磊,李刚,汪贞,李妍,张纪兵..高温纤维素降解菌的筛选及内切酶基因的克隆表达[J].农业环境科学学报,2014,(4):788-793,6.基金项目
环保公益性行业科研专项（201209036）；环保公益性行业科研专项（201309036）；中央级公益性科研院所基本科研业务专项“猪粪有机肥在有机蔬菜生产过程中的安全评价研究” （）
