玉米黄素糖基化酶在大肠杆菌中的表达与纯化

Expression and Purification of Zeaxanthin Glycosylase in E. coli

DOI：10.3969/j.issn.1673-1689.2015.08.015

中文关键词: 噬夏孢欧文氏菌 玉米黄素糖基化酶 活性

英文关键词: Erwinia uredovora, zeaxanthin glycosylase, activity

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中文摘要:

噬夏孢欧文氏菌合成类胡萝卜素涉及的一系列反应都是在相关酶的催化下完成的，其中催化玉米黄素转化为玉米黄素二葡萄糖苷的玉米黄素糖基化酶由基因crtX编码。PCR扩增出噬夏孢欧文氏菌crtX基因，构建重组质粒，转化大肠杆菌。经镍离子树脂亲和层析、DEAE-Sepharose FF离子交换层析纯化，得到了电泳纯的重组噬夏孢欧文氏菌玉米黄素糖基化酶，该重组蛋白在体外具有催化玉米黄素转化为玉米黄素二葡萄糖苷的活性。

英文摘要:

Several carotenoids are synthesized by related enzymes in Erwinia uredovora，. Zeaxanthin diglucoside is synthesized by zeaxanthin glycosylase which was encoded by gene crtX. In this study，crtX of E. uredovora was amplified by PCR and cloned into expression vector. The recombinant plasmid was then transformed into E. coli BL21 （DE3） to construct the engineering bacterium. The recombinant zeaxanthin glycosylase was purified by Ni2+ chelating affinity chromatography and a DEAE-Sepharose FF ion-exchange chromatography. The purified protein catalyze the formation of zeaxanthin diglucoside in vitro.

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