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

作　　者：孟广荣[1] 杨树林[1] 曾亮亮[1] 李新柱[1] 蔡凤[1] 

机构地区：[1]南京理工大学生物工程研究所,江苏南京210094

出　　处：《食品与生物技术学报》2006年第2期24-27,共4页Journal of Food Science and Biotechnology

基　　金：国家"十五"科技攻关专项项目(2001BA706B-18);南京市科学技术局项目(2003010145)

摘　　要：利用A¨KTA UPC-900快速蛋白液相色谱系统(FPLC)从黑曲霉发酵粉中分离纯化出内切β-葡聚糖苷酶。分离纯化后的酶比活力提高了8.1倍,回收率为7.5%。经SDS-PAGE电泳分析该内切酶的相对分子质量为26 400。酶学试验研究表明:该酶的最适反应温度为55℃,最适pH为4.8,Lineweaver-Burk法求得动力学参数,Km和Vmax分别为6.838×10-3mg/mL、2.906×10-2(mL.min)/mg。并确定了FPLC层析缓冲液的离子强度为4.8 mmol/L时分离效果达到最佳。An endo-β-glucanase was isolated and purified from a commercial preparation of Aspergillus niger by means of FPLC （AKTA UPC-900）. The specific activity of the endoglucanase increased 8.1-fold, and recovery coefficient reached 7.5 %. The molecular weight was estimated as 26 400 by SDS-PAGE. The optimal reaction temperature and pH of the endoglucanase 55 ℃ and 4. 8, respectively. The values of Km and Vmax calculated from Lineweaver-Burk plots were 6. 838 10^-3 mg/ml and 2. 906 10^-2 （ml min）/mg, respectively. The endoglucanase was purified effectively under the buffer ionic strength of 4.8 mmol/L.

关 键 词：黑曲霉 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[生物学—生物工程]