产壳聚糖酶菌株发酵条件优化及壳聚糖酶的分离纯化研究  

Study on optimal fermentation conditions of a chitosanase-producing bacterium and purification of the chitosanase

作　　者：段妍[1] 韩宝芹[2] 董文[2] 杨艳[2] 常菁[2] 刘万顺[2] 

机构地区：[1]中山大学神经与认知科技创新平台,广东广州510080 [2]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《海洋科学》2009年第1期1-7,共7页Marine Sciences

基　　金：国家"十五"科技攻关项目(2001BA708B04-07)

摘　　要：从土壤中筛选得到一株产壳聚糖酶菌株P003,对其进行了发酵条件优化。结果表明其最适发酵培养条件为:(NH4)2SO42.0%,粉末壳聚糖1.0%,葡萄糖0.1%,K2HPO40.07%,KH2PO40.03%,NaCl0.5%,MgSO4·7H2O0.05%,酵母提取物0.3%,调pH至5.0;2.0%接种量,500mL三角瓶装液量为150mL,32℃下150r/min摇床培养96h,在此条件下菌株发酵液的壳聚糖酶活力达108U/mL。采用硫酸铵分级盐析、离子交换层析和凝胶过滤层析等方法,从菌株的发酵液中分离出了分子质量为30.5ku的壳聚糖酶。A strain with chitosanase activity was isolated from soil samples. The optimal compositions of the medium for P003 strain producing chitosanase were as follows（W/F）： powder chitosan 1.0%, glucose 0.1%, （NH4）2SO42.0%, yeast extract 0.3%, K2HPO40.07%, KH2PO40.03%, NaCl 0.5%, MgSO4·7H2O 0.05%, the initial pH5.0. The optimal cultivation conditions for P003 producing chitosanase were 500 mL flask containing 150 mL medium, 2.0% （V/V） amount of inoculation , 32℃ incubating temperature and shaking cultivation at 150 r/min for 96 h. The chitosanase activity of the fermented broth is 108 U/mL when P003 was incubated under aforementioned conditions. The chitosanase of the fermented broth was extracted by ammonium sulfate precipitation, and purified by ion-exchange chromatography and gel filtration, and the molecular weight of the chitosanase was estimated to be 30.5 ku.

关 键 词：壳聚糖酶 发酵条件 分离纯化 

分 类 号：Q814.1[生物学—生物工程]