Manipulating the stereoselectivity of the thermostable Baeyer–Villiger monooxygenase TmCHMO by directed evolution†,Organic & Biomolecular Chemistry

当前位置： X-MOL 学术 › Org. Biomol. Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Manipulating the stereoselectivity of the thermostable Baeyer–Villiger monooxygenase TmCHMO by directed evolution†
Organic & Biomolecular Chemistry ( IF 2.9 ) Pub Date : 2017-11-13 00:00:00 , DOI: 10.1039/c7ob02692g
Guangyue Li _{1,

2,

3,

4,

5} , Maximilian J. L. J. Fürst _{6,

7,

8,

9} , Hamid Reza Mansouri _{10,

11,

12,

13} , Anna K. Ressmann _{10,

11,

12,

13} , Adriana Ilie _{1,

2,

3,

4,

5} , Florian Rudroff _{10,

11,

12,

13} , Marko D. Mihovilovic _{10,

11,

12,

13} , Marco W. Fraaije _{6,

7,

8,

9} , Manfred T. Reetz _{1,

2,

3,

4,

5}

Affiliation

Baeyer–Villiger monooxygenases (BVMOs) and evolved mutants have been shown to be excellent biocatalysts in many stereoselective Baeyer–Villiger transformations, but industrial applications are rare which is partly due to the insufficient thermostability of BVMOs under operating conditions. In the present study, the substrate scope of the recently discovered thermally stable BVMO, TmCHMO from Thermocrispum municipale, was studied. This revealed that the wild-type (WT) enzyme catalyzes the oxidation of a variety of structurally different ketones with notable activity and enantioselectivity, including the desymmetrization of 4-methylcyclohexanone (99% ee, S). In order to induce the reversal of enantioselectivity of this reaction as well as the transformations of other substrates, directed evolution based on iterative saturation mutagenesis (ISM) was applied, leading to (R)-selectivity (94% ee) without affecting the thermostability of the biocatalyst.

中文翻译：

通过定向进化来操纵热稳定的Baeyer-Villiger单加氧酶TmCHMO的立体选择性†

在许多立体选择性的Baeyer-Villiger转化中，Baeyer-Villiger单加氧酶（BVMOs）和进化的突变体已被证明是出色的生物催化剂，但是工业应用很少见，部分原因是在操作条件下BVMOs的热稳定性不足。在本研究中，研究了最近发现的热稳定BVMO，即来自Thermocrispum Municipale的TmCHMO的底物范围。这表明野生型（WT）酶催化具有显着活性和对映选择性的多种结构不同的酮的氧化，包括4-甲基环己酮（99％ee，S）。为了诱导该反应的对映选择性以及其他底物的转化的逆转，应用了基于迭代饱和诱变（ISM）的定向进化，从而导致（R）选择性（94％ee），而不会影响反应的热稳定性。生物催化剂。

更新日期：2017-11-13

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11