产纳豆激酶菌株Bacillus sp.ZLK08的分离及酶纯化  

Isolation of Nattokinase-Producing Bacillus sp.ZLK08 and Enzyme Purification

作　　者：李蓓[1] 龙梅[1] 郭放[1] 何雪梅[1] 邹立扣[1] 罗燕[1] 

机构地区：[1]四川农业大学都江堰校区微生物学实验室,四川都江堰611830

出　　处：《微生物学杂志》2012年第2期98-101,共4页Journal of Microbiology

基　　金：四川农业大学"双支"计划资助

摘　　要：获得稳定产纳豆激酶菌株,为高酶活纳豆激酶产生菌的改造奠定基础。取各来源纳豆,用平板梯度稀释法分离菌株,测定菌株酶活,利用16S rDNA鉴定产酶菌株,凝胶过滤法纯化纳豆激酶并SDS-PAGE检测分析。成功获得稳定产酶芽胞杆菌Bacillus sp.ZLK08,16S rDNA分析表明其与GenBank中序列同源率达到99%,液体发酵表明酶活达到2.5 FU/mL,经纯化,SDS-PAGE表明纳豆激酶分子量为28.46 ku。分离出的Ba-cillus sp.ZLK08菌株能稳定产纳豆激酶且具有较高酶活。In order to lay a foundation to reform nattokinase-producing strain,to obtain a stable nattokinase-producing strain,nattoes collected from various sources were isolated using plate gradient dilution method,and adopted 16S rDNA to characterize the enzyme-producing strain,finally purified the nattokinase with gel filtration and tested,analyzed with SDS-PAGE.A stable nattokinase-producing strain Bacillus sp.ZLK08 was successfully obtained.16S rDNA analysis indicated that the sequence homologous rate was as high as 99% with that in GenBank.The enzyme activity by liquid fermentation of the strain Bacillus sp.ZLK08 was as high as 2.5 FU/mL.SDS-PAGE indicated that molecular mass of the nattokinase was 28.46 ku after it was purified.Therefore,the isolated Bacillus sp.strain ZLK08 could stably produce nattokinase possessing fairly high enzyme activity.

关 键 词：纳豆激酶 芽胞杆菌 纯化 

分 类 号：Q93[生物学—微生物学] Q814