绿色木霉MJ1产内切β-葡聚糖苷酶的分离纯化及酶学性质  

Purification and Properties of an Endo-β-glucanase from Trichoderma viride MJ1

作　　者：杨树林[1] 孟广荣[1] 李校堃[2] 曾亮亮[1] 富国平[1] 

机构地区：[1]南京理工大学生物工程研究所,南京210094 [2]吉林农业大学生物反应器与药物开发教育部工程中心,长春130118

出　　处：《微生物学通报》2006年第5期75-79,共5页Microbiology China

基　　金：国家高技术研究发展计划项目("863"项目)(No.2004AA2Z3C60);国家十五科技攻关项目(No.2001BA706B-18)

摘　　要：利用KTAUPC-900快速蛋白液相色谱系统(FPLC)从绿色木霉MJ1固体发酵产物中分离纯化出内切β-葡聚糖苷酶。分离纯化后酶的比活力提高了28·6倍,回收率为19·7%。SDS-PAGE后经BIO-RAD凝胶成像系统分析该内切酶的分子量为64·7kD。酶学试验研究表明:该酶的最适反应温度53℃,最适pH为4·2,Lineweaver-Burk法求得动力学参数,Km和Vmax分别为1·230×10-2g/mL、2·396×10-2mg/(mL·min)。并确定了FPLC层析缓冲液的离子强度为2·2mmol/L时分离效果达到最佳。An endo-β-glucanase was isolated and purified from the leavening of Trichoderma viride called MJ1 by means of FPLC （AKTA UPC-900） . The specific activity of the endoglucanase increased 28.6 fold, and coefficient of recovery was 19.7%. The molecular mass was 64. 7kD determined by gel imaging system after SDS-PAGE. The optimal reaction temperature of the endoglucanase was 53℃, and the optimal pH was 4.2. The values of Km and Vmax calculated from Lineweaver-Burk plots were 1. 230 × 10^-2g/mL and 2. 396 × 10^-2 mg/ （ mL · min） respectively. The endoglucanase was purified effectively when the ionic strength of the buffer was 2.2mmoL/L.

关 键 词：绿色木霉 纤维素酶 内切Β-葡聚糖苷酶 分离纯化 酶学性质 动力学 

分 类 号：Q814.1[生物学—生物工程]