海洋微生物几丁质酶分离纯化及其抗真菌活性  

Separation and Purification of a Chitinase from Marine Microbe and Determination of the Antifungal Activity

作　　者：丁存宝[1] 刘海燕[1] 贾长虹[1] 张俊杰[1] 

机构地区：[1]河北理工大学化工与生物技术学院唐山市海洋资源综合利用重点实验室,河北唐山063009

出　　处：《微生物学杂志》2009年第4期67-70,共4页Journal of Microbiology

基　　金：唐山市科技局项目(04364001B-11)

摘　　要：以实验室筛选的海洋产几丁质酶短芽胞杆菌属(Bacillus brevis sp.)菌株Bsp1,经往复式摇床振荡培养96 h后,发酵液先后采取了75%的硫酸铵盐析、透析、几丁质亲和层析、SDS-PAGE等方法对几丁质粗酶液进行分离纯化和鉴定。几丁质亲和层析一步纯化后,经过SDS-PAGE电泳测定该酶的分子量为23 ku,其比活力为86.65,纯化倍数为1.707、产率为32.1%。纯化的几丁质酶能抑制病原真菌的生长,对病原真菌的拮抗作用具有广谱性。同时研究了几丁质酶的稳定性,以胶态几丁质为底物,分离的几丁质酶在pH7.5,55.0℃左右具有最大酶活性;Zn2+、Cu2+和Hg2+能强烈抑制几丁质酶活性;Ni+和EDTA抑制20%～40%;然而5mmol/L Co2+可以使几丁质酶活性提高1.4倍;Mg2+、Ca2+等也能使酶活性增加。After 30 qC cultivation for 96 hours, the chitinase from a Bacillus brevis sp. named Bspl was extracted by 75% ammonium sulfate precipitation and then dialysed. The obtained ehitinase was purified by chitin.based affinity chromatography. The purified chitinase showed a single band with a molecular weight of 23 ku in gel after SDS-PAGE. The chitinase displayed a 86.65 specific activity, 1. 707 purified fold, and a 32. 1% yield. The crude ehitinase showed the inhibition to a wide-range of pathogenic fungi. The purified ehitinase has maximal activity at pH7.5 and 55.0 ℃ , but Zn^2＋ , Cu^＋ and Hg^2＋ inhibited the activity while 5 mmol/L Co^2＋ increased its activity.

关 键 词：几丁质酶 分离纯化 抗真菌活性 

分 类 号：S917.1[农业科学—水产科学]