Designing new Baeyer-Villiger monooxygenases using restricted CASTing
Designing new Baeyer-Villiger monooxygenases using restricted CASTing
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DOI：
10.1021/jo0613636
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发表时间：
2006-10-27
影响因子：
3.6
通讯作者：
Reetz, Manfred T.
中科院分区：
文献类型：
--
作者：
Clouthier, Christopher M.;Kayser, Margaret M.;Reetz, Manfred T.
本文概述了环戊酮单加氧酶（CPMO）第一次微进化的设计和实现。所描述的方法是一种相对便宜和快速的方法来获得具有所需特性的突变酶。几个对映体选择性增强的成功突变体被鉴定出来。例如，突变体催化的4-甲氧基环己酮氧化使相应的内酯具有92%的多余量（ee），而野生型环己酮单加氧酶（WT-CHMO）的多余量为46%。迷你进化的原始设计和突变体的后续评估可以为活性位点的构建和动态提供有价值的见解，并可以在假定的活性位点内提出其他催化有益的突变。
This paper outlines the design and execution of the first mini-evolution of cyclopentanone monooxygenase (CPMO). The methodology described is a relatively inexpensive and rapid way to obtain mutant enzymes with the desired characteristics. Several successful mutants with enhanced enantioselectivities were identified. For example, mutant-catalyzed oxidation of 4-methoxycyclohexanone gave the corresponding lactone with 92% entantiometric excess (ee) compared to the 46% ee achieved with wild-type cyclohexanone monoxygenase (WT-CHMO). The original design of the mini-evolution and the following evaluation of mutants can provide valuable insights into the active site's construction and dynamics and can suggest other catalytically profitable mutations within the putative active site.