Lactone-Bound Structures of Cyclohexanone Monooxygenase Provide Insight into the Stereochemistry of Catalysis

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• 作者：Brahm J. Yachnin ; Michelle B. McEvoy ; Roderick J. D. MacCuish ; Krista L. Morley ; Peter C. K. Lau ; Albert M. Berghuis
• 刊名：ACS Chemical Biology
• 出版年：2014
• 出版时间：December 19, 2014
• 年：2014
• 卷：9
• 期：12
• 页码：2843-2851
• 全文大小：320K
• ISSN：1554-8937

文摘

The Baeyer鈥揤illiger monooxygenases (BVMOs) are microbial enzymes that catalyze the synthetically useful Baeyer鈥揤illiger oxidation reaction. The available BVMO crystal structures all lack a substrate or product bound in a position that would determine the substrate specificity and stereospecificity of the enzyme. Here, we report two crystal structures of cyclohexanone monooxygenase (CHMO) with its product, 蔚-caprolactone, bound: the CHMO_Tight and CHMO_Loose structures. The CHMO_Tight structure represents the enzyme state in which substrate acceptance and stereospecificity is determined, providing a foundation for engineering BVMOs with altered substrate spectra and/or stereospecificity. The CHMO_Loose structure is the first structure where the product is solvent accessible. This structure represents the enzyme state upon binding and release of the substrate and product. In addition, the role of the invariant Arg329 in chaperoning the substrate/product during the catalytic cycle is highlighted. Overall, these data provide a structural framework for the engineering of BVMOs with altered substrate spectra and/or stereospecificity.