Lactone-bound structures of cyclohexanone monooxygenase provide insight into the stereochemistry of catalysis
| Autor: | Michelle B. McEvoy, Brahm J. Yachnin, Albert M. Berghuis, Krista Morley, Peter C. K. Lau, Roderick J. D. MacCuish |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular crystal structure Stereochemistry oxidation Gene Expression Crystallography X-Ray Protein Engineering 010402 general chemistry 01 natural sciences Biochemistry Substrate Specificity Catalysis Lactones Stereospecificity Bacterial Proteins stereospecificity Oxidoreductase polycaprolactone protein conformation protein cross linking Rhodococcus Binding site Caproates chemistry.chemical_classification Binding Sites conformational transition catalysis 010405 organic chemistry Chemistry binding site microbial enzyme stereochemistry Substrate (chemistry) Stereoisomerism General Medicine Protein engineering unspecific monooxygenase 0104 chemical sciences enzyme structure enzyme Catalytic cycle Biocatalysis lactone Mutation Oxygenases Molecular Medicine cyclohexanone Protein Binding |
| Popis: | The Baeyer-Villiger monooxygenases (BVMOs) are microbial enzymes that catalyze the synthetically useful Baeyer-Villiger 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, ε-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. |
| Databáze: | OpenAIRE |
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