Molecular Enzymology Group, University of Groningen , Nijenborgh 4, 9747AG, Groningen, The Netherlands.
Department of Biology and Biotechnology, University of Pavia , Via Ferrata 1, 27100, Pavia, Italy.
J Am Chem Soc. 2017 Jan 18;139(2):627-630. doi: 10.1021/jacs.6b12246. Epub 2017 Jan 3.
Regio- and stereoselective Baeyer-Villiger oxidations are difficult to achieve by classical chemical means, particularly when large, functionalized molecules are to be converted. Biocatalysis using flavin-containing Baeyer-Villiger monooxygenases (BVMOs) is a well-established tool to address these challenges, but known BVMOs have shortcomings in either stability or substrate selectivity. We characterized a novel BVMO from the thermophilic fungus Thermothelomyces thermophila, determined its three-dimensional structure, and demonstrated its use as a promising biocatalyst. This fungal enzyme displays excellent enantioselectivity, acts on various ketones, and is particularly active on polycyclic molecules. Most notably we observed that the enzyme can perform oxidations on both the A and D ring when converting steroids. These functional properties can be linked to unique structural features, which identify enzymes acting on bulky substrates as a distinct subgroup of the BVMO class.
区域和立体选择性 Baeyer-Villiger 氧化反应很难通过传统的化学方法实现，特别是在需要转化大的、功能化分子时。使用含有黄素的 Baeyer-Villiger 单加氧酶 (BVMO) 的生物催化是解决这些挑战的一种成熟工具，但已知的 BVMO 在稳定性或底物选择性方面存在缺点。我们从嗜热真菌 Thermothelomyces thermophila 中鉴定出一种新型的 BVMO，测定了其三维结构，并证明其可用作有前途的生物催化剂。这种真菌酶表现出优异的对映选择性，可作用于各种酮，并且对多环分子特别活跃。值得注意的是，我们观察到该酶在转化甾体时可以同时对 A 环和 D 环进行氧化。这些功能特性可以与独特的结构特征相关联，这些特征将作用于大体积底物的酶鉴定为 BVMO 类的一个独特亚组。
