文献类型：期刊文献

中文题名：大肠杆菌核苷磷酸化酶的重组表达和活性

英文题名：Cloning,Expression and Activity Analysis of Nucleoside Phosphorylases Genes from Escherichia coli K12

作者：谭黎[1];欧阳立明[1];丁庆豹[1];欧伶[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2008

卷号：34

期号：5

起止页码：660

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：20084811740418);Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：大肠杆菌;嘌呤核苷磷酸化酶;尿苷磷酸化酶;胸苷磷酸化酶;表达

外文关键词：Escherichia coli ; pyrimidine nucleoside phosphorylase; uridine phosphorylase; thymidine phosphorylase ; expression

摘要：将大肠杆菌K-12菌株来源的嘌呤核苷磷酸化酶、尿苷磷酸化酶和胸苷磷酸化酶基因分别克隆到pET-11a载体并转化大肠杆菌BL21(DE3)宿主菌表达,通过SDS-PAGE分析和酶的活性测定,重组菌诱导表达后目的蛋白的表达量占菌体总蛋白的37%以上,与产核苷磷酸化酶的野生菌比较,酶的活性也有显著提高。在特异反应条件下,构建的工程菌可以用来高效催化核苷的转糖基反应。
The genes encoding pyrimidine nucleoside phosphorylase, uridine phosphorrylase and thy- midine phosphorylase from Escherichia coli K12 were cloned respectively into expression vector pET-11a. The recombinant plasmids were then transformed into the strain E. coli BL21（DE3）. As a result the enzymes were highly expressed after induction with IPTG. The expression product was analyzed with SDS- PAGE varifying that target proteins were expressed in soluble form and the amount accounts for more than 37% of total protein in the host. Compared with the wild strains which also produce nucleoside phosphorylase well, the enzyme activity of three recombinant strains is evidently improved. In the specific reaction conditions, the new constructed strains can be used to transfer ribose or deoxyribose from a nucleoside to a new base efficiently.