利用Native制备电泳分离来源于Rhizopus microsporus var. chinensis的葡萄糖淀粉酶同工酶  

Isolation of Two Glucoamylase Isoenzymes from Rhizopus microsporus var. chinensis by Native Prep Cell

作　　者：李彧娜[1,2] 石贵阳[1,2] 王武[1,2] 王正祥[1,2] 

机构地区：[1]江南大学生物工程学院工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院生物资源与生物能源研究中心,江苏无锡214122

出　　处：《食品与发酵工业》2010年第1期1-4,共4页Food and Fermentation Industries

基　　金：国家高技术研究发展计划"863"项目(No.2006AA020204)资助

摘　　要：将Native制备电泳应用于Rhizopus microsporus var. chinensis中2个葡萄糖淀粉酶同工酶的分离,考察了电泳缓冲系统、凝胶浓度和凝胶长度对分离效果的影响。结果表明:先使用Ornstein-Davis系统分离同工酶,再使用0.02mol/L,pH6.2的NaAc.CH3COOH缓冲液洗脱,在长度为6cm的7%的Native电泳胶上,2个性质相近仅电泳迁移率略有差别的同工酶能有效分离。分离回收的蛋白质可保持其活力,便于后续研究。Two isoenzymes of glucoamylase secreted by strain of Rhizopus microsporus var. chinensis were separated from each other by Native Prep Cell. Three factors including electrophoretic buffer system, gel composition and gel height were investigated. Results indicated when experimental conditions were as follows： Ornstein-Davis electrophoretic buffer system for ionophortic separation, 0.02 mol/L, pH 6.2 NaAc · CH3COOH washing system for protein elution, 7% gel composition and 6cm gel height for Native PAGE, two isoenzymes with very similar characters could be separated by their different electrophoretic mobility. Moreover, the two isoenzymes maintained their activities after electrophoresis, which contributed to the characterization of the properties of purified proteins.

关 键 词：制备电泳 RHIZOPUS microsporus var.chinensis 葡萄糖淀粉酶 同工酶 

分 类 号：Q814[生物学—生物工程]