猴头菌UDP-葡萄糖-4-差向异构酶基因克隆表达及酶学性质  

Expression and enzymatic property of a UDP-glucose-4-epimerase gene cloned from Hericium erinaceus

作　　者：任鄄宝 邹根 张忠[2] 陈万超[2] 吴迪[2] 张赫男[2] 龚明[2] 杨焱[2] REN Juan-Bao;ZOU Gen;ZHANG Zhong;CHEN Wan-Chao;WU Di;ZHANG He-Nan;GONG Ming;YANG Yan(College of Food Science,Shanghai Ocean University,Shanghai 201306,China;National Engineering Research Center of Edible Fungi,Key Laboratory of Resources and Utilization of Edible Fungi(South),Ministry of Agriculture,Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]国家食用菌工程技术研究中心农业部南方食用菌资源利用重点实验室上海市农业科学院食用菌研究所,上海201403

出　　处：《菌物学报》2021年第9期2330-2340,共11页Mycosystema

基　　金：上海市科委重点研发项目(18391903500);上海市科技兴农重点攻关项目(沪农科种字[2016]第1-12号);上海市领军人才项目(2018)。

摘　　要：通过实验室前期对诱变菌株猴头菌321的多组学分析结果,获得了一个多糖合成过程中起关键作用的UDP-葡萄糖-4-差向异构酶基因(UDP-glucose-4-epimerase,UGE),并在大肠杆菌E. coli BL21(DE3)中进行了异源表达。通过筛选最优的目的蛋白诱导表达条件后,通过镍柱亲和层析纯化后获得高纯度目的蛋白,并对目的蛋白进行了酶学性质的研究,明确了其生物学性质和动力学参数,为其开发利用提供理论参考。A UDP-glucose-4-epimerase(UGE) gene, which plays a key role in the synthesis of polysaccharides, was obtained through multi-omics analysis of the mutated strain Hericium erinaceus 321, and was heterologously expressed in E. coli BL21(DE3). The inductive expression conditions were optimized. The high purity target protein was obtained after purification by nickel column affinity chromatography. The enzymatic properties of the protein were studied, and the optimal activity temperature and pH were determined. The biological properties and kinetic parameters of the protein were defined. The results provided theoretical reference for development and utilization of the gene.

关 键 词：基因克隆 诱导表达 蛋白纯化 酶学性质 

分 类 号：Q78[生物学—分子生物学]