壳聚糖酶高产菌株的筛选、鉴定及发酵条件的研究  

Screening,Identification and Optimization of Fermentation Conditions of A High-yield Chitosanase-producing Strain of Bacillus

作　　者：孙玉英[1] 张继泉[2] 王淑军[1] 

机构地区：[1]淮海工学院海洋学院,江苏连云港222005 [2]中国科学院海洋研究所,山东青岛266071

出　　处：《食品科学》2009年第15期164-168,共5页Food Science

基　　金：淮海工学院引进人才科研启动项目(KQ03104);淮海工学院自然科学基金项目(Z2008041)

摘　　要：从连云港海滩晒虾蟹壳的泥土里筛选出一株产壳聚糖酶能力较高的菌株S-1(水解圈直径和菌落直径的比值高达12.5)。根据其形态特征、生理生化以及16SrDNA鉴定,初步确定该菌为芽孢杆菌属(Bacillus);采用单因子试验方法,确定该菌的最佳培养基配方为(g/L):虾蟹壳粉15、硝酸铵10、酵母提取物3、K2HPO4·3H2O1.4、NaCl5、MgSO4·7H2O1.4、葡萄糖1.0;利用正交试验进行发酵条件的优化,确定最适宜发酵条件为初始pH6.0,32℃,用500ml三角瓶装液量为100ml,发酵时间为72h,最终该菌的酶活力由复筛的2.13U/ml提高到6.4U/ml,从而使该菌株具有工业化应用的潜力。Twenty chitosanase-producing bacterium strains were isolated from Lianyungang seaside. Among them strain S-1 had the highest chitosanase-producing capacity and was identified as Bacillus sp. based on its morphological, physiological and biochemical properties as well as 16S rDNA sequence analysis. Single-factor method and orthogonal array design were adopted to optimize the medium composition and fermentation parameters to maximize chitosanase activity, respectively. The optimal medium for enzyme production consisted of 15 g/L shrimp- and crab-shell powder, 10 g/L NH4NO3, 3 g/L yeast extract, 1.4 g/L K2HPO4 · 3H2O, 5 g/L NaCI, 1.4 g/L MgSO4 · 7H2O and 1 g/L glucose at an initial pH of 6.0 and the optimal fdling volume was 100 ml in 500 ml flask. After cultivation at 32 ℃ for 72 h, the maximal chitosanase activity was observed to be 6.4 U/ml. This strain has promising potential in the industrial production of chitosanase.

关 键 词：壳聚糖酶 筛选 鉴定 发酵优化 芽孢杆菌 

分 类 号：Q53[生物学—生物化学]