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首页> 外文会议>International citrus congress >Functional Expression of Phytoene Synthase Gene from Cara Cara Navel Orange
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Functional Expression of Phytoene Synthase Gene from Cara Cara Navel Orange

机译：Cara Cara Navel橙的植物合酶基因的功能表达

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摘要

Citrus is a rich source of carotenoids. Phytoene synthase (PSY, EC: 2. 5. 1. 32 ), an important rate-limiting enzyme, converts geranylgeranyl diphosphate (GGPP) to phytoene in the first committed step of the carotenoid biosynthetic pathway. To further characterize PSY we investigated catalytic activity of CsPSY from Cara Cara navel orange by heterologous expression in E. coli strains. The gene was cloned into pET-28a (+)vector system controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CsPSY protein is normally expressed in E. coli. In vivo complementation and HPLC analysis revealed that phytoene was produced in recombinant E. coli, which indicated that CsPSY was functionally expressed in E. coli and catalyze a head-to-head condensation of two molecules of GGPP to form phytoene. This study suggests that CsPSY can generally complement the corresponding bacterial enzyme and could be used as a means of engineering bacteria and plants for the production of carotenoids.
机译：柑桔是一种富含类胡萝卜素的来源。八氢番茄红素合酶（PSY，EC：2. 5. 1. 32），一个重要的限速酶，香叶基香叶基变换二磷酸（GGPP），以八氢番茄红素在类胡萝卜素生物合成途径的第一个关键步骤。为了进一步表征PSY我们通过在大肠杆菌菌株异源表达研究从红肉脐橙CsPSY的催化活性。的基因被克隆到由T7启动子控制的pET-28a中（+）载体系统。 SDS聚丙烯酰胺凝胶电泳表明，CsPSY蛋白在大肠杆菌中正常表达。活体内互补和HPLC分析显示，八氢番茄红素在重组大肠杆菌，表明CsPSY在大肠杆菌中功能性表达，并催化GGPP的两个分子的头 - 头缩合以形成八氢番茄红素生产。这项研究表明，CsPSY通常可以补充相应的细菌酶和可以被用作工程细菌和植物用于生产类胡萝卜素的一种手段。

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