一株嗜热厌氧菌的分离及其耐高温淀粉酶基因的克隆和表达  

Isolation and characterization of Thermoanaerobacter sp.185y2,clone and expression of its thermophilic α-amylase gene

作　　者：赵子如[1] 梅琴[1] 高雅英[1] 虞方伯[1] 沈标[1] 

机构地区：[1]南京农业大学生命科学学院/农业部农业环境微生物工程重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2007年第1期55-60,共6页Journal of Nanjing Agricultural University

基　　金：科技部农业微生物菌种资源整理整合及共享项目(2005DKA21201-11)

摘　　要：从云南腾冲眼镜泉中分离到1株嗜热厌氧的杆状细菌185y2。根据其生理生化特征和16S rDNA序列同源性分析,将该菌株鉴定为嗜热厌氧杆菌属Therm oanaerobactersp.。该菌的最适生长pH值和温度分别为7.0和70℃,具有水解淀粉的能力。从185y2的基因组DNA中扩增出高温淀粉酶结构基因,序列分析表明,它与中温菌的淀粉酶基因亲缘关系较远。克隆其耐高温淀粉酶基因,并构建重组表达载体,最终在E.coliBL21中得到了高效表达。A thermophilc anaerobic bacterium 185y2 was isolated from a hot spring in Tengchong, Yunnan Province. This strain was identified as Thermoanaerobacter sp. according to its physiological and biochemical characteristics and phylogenetic analysis based on the 16S rDNA sequence. The optimum pH and temperature for the growth of the isolate were 7. 0 and 70 ℃ respectively. This isolate had the ability to hydrolyse the starch. The gene encoding amylase was amplified from the genome DNA of 185y2 by PCR, which was lower similarity to the mesophic amylase gene. The recombinant plasmid pET-amy was constructed and amylase gene was expressed in E. coli BL21.

关 键 词：嗜热厌氧菌 分离 鉴定 高温淀粉酶 

分 类 号：Q939.99[生物学—微生物学]