熊果苷合成酶固定化条件研究  

Immobilization conditions for arbutin synthetase

作　　者：李皓[1] 盛希群 刘静[1] 钟星[1] LI Hao;SHENG Xi-qun;LIU Jing;ZHONG Xing(Department of Biological and Chemical Engineering,Zhixing College of Hubei University,Wuhan 430011,China)

机构地区：[1]湖北大学知行学院生物与化学工程学院,武汉430011

出　　处：《湖北农业科学》2021年第2期85-89,共5页Hubei Agricultural Sciences

摘　　要：将转化至Escherichia coli BL21 Gold(DE3)中aglA基因的质粒表达葡萄糖苷酶用以催化生产α-熊果苷。再利用超声波破碎技术和镍离子层析柱提取并纯化转葡萄糖苷酶,并将其用海藻酸钠-明胶混合固定,使其可重复使用。确定最佳的固定化条件,即将3.0%海藻酸钠和3.0%明胶配制溶解于pH 7的磷酸氢二钠-磷酸二氢钠缓冲溶液的混合胶体,在高度45 cm左右进行滴定4℃预冷的300 g/L氯化钙溶液并固定化20 min。冲洗后进行50 min的-20℃钙化操作,获得直径为3.97 mm、质量为0.311 g的固定化珠。The constructed plasmid containing aglA gene was transformed into Escherichia coli BL21 Gold(DE3)to induce its suc⁃cessful expression,and the transglucosidase was synthesized to catalyze the production ofα-arbutin.Then the transglucosidase was ex⁃tracted and purified by ultrasonic crushing technique and nickel ion chromatography column and fixed by sodium alginate-gelatin mix⁃ture so that it could be reused.The optimum immobilization conditions were as follows:A mixture of colloid dissolved in pH 7 with 3.0%sodium alginate and 3.0%gelatin was prepared.The solution was titrated at a height of about 45 cm and pre-cooled at 4℃by 300 g/L calcium chloride solution and immobilized for 20 min.After washing,calcification was performed at-20℃for 50 min to ob⁃tain immobilized beads with a diameter of 3.97 mm and a mass of 0.311 g.

关 键 词：诱导表达 α-熊果苷 转葡萄糖苷酶 固定化 

分 类 号：TQ460.1[化学工程—制药化工] TQ658