Directed evolution of squalene synthase for dehydrosqualene biosynthesis
Directed evolution of squalene synthase for dehydrosqualene biosynthesis
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DOI：
10.1016/j.febslet.2014.07.028
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发表时间：
2014-09-17
期刊：
影响因子：
3.5
通讯作者：
Umeno, Daisuke
中科院分区：
文献类型：
--
作者：
Furubayashi, Maiko;Li, Ling;Umeno, Daisuke
角鲨烯合酶（SQS）催化多种生物体中甾醇/类藿烷生物合成的第一步。长期以来，人们已经认识到SQS与类胡萝卜素脱氢酶有着共同的祖先，但不知道这些酶如何选择性地产生它们自己的产物。在这项研究中，SQS从酵母，人类和细菌分别进行定向进化的C-30类胡萝卜素骨架，脱氢角鲨烯的生产。这是通过用Pantoea ananatis八氢番茄红素去饱和酶进行高通量筛选来实现的，该酶可以选择性地将脱氢角鲨烯转化为黄色类胡萝卜素色素。对所产生的突变体的遗传分析揭示了各种可以有效地将SQS转化为“脱氢角鲨烯合酶”的突变。所有这些突变都聚集在被认为对NADPH结合很重要的残基周围。(C)2014年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Squalene synthase (SQS) catalyzes the first step of sterol/hopanoid biosynthesis in various organisms. It has been long recognized that SQSs share a common ancestor with carotenoid synthases, but it is not known how these enzymes selectively produce their own product. In this study, SQSs from yeast, human, and bacteria were independently subjected to directed evolution for the production of the C-30 carotenoid backbone, dehydrosqualene. This was accomplished via high-throughput screening with Pantoea ananatis phytoene desaturase, which can selectively convert dehydrosqualene into yellow carotenoid pigments. Genetic analysis of the resultant mutants revealed various mutations that could effectively convert SQS into a "dehydrosqualene synthase.'' All of these mutations are clustered around the residues that have been proposed to be important for NADPH binding. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.