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首页> 外文期刊>Combinatorial Chemistry _High Throughput Screening >High-Throughput Selection System for Assessing the Activity of Epoxide Hydrolases
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High-Throughput Selection System for Assessing the Activity of Epoxide Hydrolases

机译：用于评估环氧水解酶活性的高通量选择系统

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摘要

Crucial to the success of directed evolution of enantioselective enzymes for use as catalysts in synthetic organic chemistry is the availability of high-throughput assays for determining the enantiopurity of thousands of samples. Although several such ee-assays are available, they entail time and effort, which means that pre-tests for activity have been developed to eliminate non-active mutants prior to ee-screening. Pre-selection systems may be even more efficient and simple to perform. In the present paper an efficient pre-selection test for assessing the activity of epoxide hydrolases has been developed. The bacterial (E. coli) growth on agar plates is shown to be directly related to the presence of active epoxide hydrolases which catalyze the detoxicating hydrolysis of the epoxide substrates. Visual inspection of agar plates is all that is necessary to identify positive (active) hits in large libraries of mutant epoxide hydrolases.
机译：在合成有机化学中用作催化剂的对映选择性酶的定向进化成功与否的关键在于，用于确定成千上万个样品的对映体纯度的高通量分析方法的可用性。尽管有几种此类ee检测方法可用，但它们需要时间和精力，这意味着已经开展了活性预测试以在ee筛选之前消除非活性突变体。预选系统甚至可能更加高效且易于执行。在本文中，已经开发了用于评估环氧化物水解酶活性的有效预选测试。琼脂平板上细菌（大肠杆菌）的生长与活性环氧化物水解酶的存在直接相关，该酶催化环氧化物底物的解毒水解。在大型的突变型环​​氧水解酶文库中，对琼脂平板进行目测检查是确定阳性（有效）命中所必需的。

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