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化学
生物化学
选择(遗传算法)
组合化学
羟基苯甲酸
计算生物学
立体化学
酶
生物
计算机科学
有机化学
抗氧化剂
人工智能
作者
Sarah Maxel,Derek Aspacio,Edward J. King,Linyue Zhang,Ana Paula Acosta,Han Li
标识
DOI:10.1021/acscatal.0c01892
摘要
We report an aerobic, growth-based selection platform founded on NADP(H) redox balance restoration in Escherichia coli, and we demonstrate its application in the high-throughput evolution of an oxygenase. A single round of selection followed by a facile growth assay enabled Pseudomonas aeruginosa 4-hydroxybenzoate hydroxylase (PobA) to efficiently hydroxylate both 4-hydroxybenzoic acid (4-HBA) and 3,4-dihydroxybenzoic acid (3,4-DHBA), two consecutive steps in gallic acid biosynthesis. Structural modeling suggests precise reorganization of active site hydrogen bond network, which is difficult to obtain without deep navigation of combinatorial sequence space. We envision universal application of this selection platform in engineering NADPH-dependent oxidoreductases.
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