一株产耐热普鲁兰酶菌株Anoxybacillus sp. LM14-2分离鉴定及酶学性质研究  

Study of a Novel Thermostable Pullulanase Producing Strain Anoxybacillus sp. LM14-2

作　　者：孙劭靖[1,2] 路福平[1] 姜楠 李丽 徐健勇 曹慕琛[1,2] 宋诙 

机构地区：[1]天津科技大学生物工程学院工业微生物教育部重点实验室,天津300457 [2]中科院天津生物技术研究所基因工程与微生物应用技术实验室,天津300308

出　　处：《生物技术通报》2011年第9期136-141,共6页Biotechnology Bulletin

基　　金：中国科学院知识创新工程重要方向项目(KSCXZ-YW-G-075-21)

摘　　要：从云南轮马热泉下游淤泥中筛选得到了一株产耐热普鲁兰酶菌株LM14-2。根据形态特征及16S rRNA序列同源性分析,初步判定为Anoxybacillus sp.LM14-2。该菌株发酵上清液中有耐热普鲁兰酶积累,其反应最适pH值为6.0,最适温度为70℃。利用染色体步移技术获得了完整的普鲁兰酶编码基因(HQ660582),经序列相似性进一步分析,确定该蛋白与I型普鲁兰酶保守区b相吻合。通常的普鲁兰酶在高温下很快失活,难以满足淀粉加工,洗涤剂等相关工业的需求,而该新型的耐热普鲁兰酶的作用温度广泛,热稳定性较好,65℃保温55 h后达到其半衰期,具有广阔的开发应用前景。A novel strain LM14-2 producing thermostable pullulanase was recovered from Lunma hot spring of Yunnan Province of China.According to colony morphology and 16S rRNA blasting,this strain was named Anoxybacillus sp.LM14-2.The thermostable pullulanase with optimum pH6.0 and optimum temperature 70℃was accumulated in zymotic fluid of the Anoxybacillus sp.LM14-2.The corresponding pullulanase gene（GenBank No.HQ660582）were obtained through gene walking.The structural domain of the pullulanase dovetailed well with that of pullulanaseⅠaccording to the analysis of sequence similarities.The pullulanase previously discovered inactivates under high temperature,which was difficult to satisfy with biotechnological applications such as sugar making,detergent,etc.However,the thermostable pullulanase produced by the novel Anoxybacillus sp.LM14-2 which features wide temperature,excellent thermal stability and a half-life as long as 55 h at 65℃,and performances widely industrial application prospect.

关 键 词：普鲁兰酶 筛选 16S RRNA 基因步移 ANOXYBACILLUS sp. 

分 类 号：TQ925[轻工技术与工程—发酵工程]