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

Yachnin, Brahm J.; McEvoy, Michelle B.; MacCuish, Roderick; Morley, Krista L.; Lau, Peter C. K.; Berghuis, Albert M.

doi:10.1021/cb500442e

Cited by 39 publications

(71 citation statements)

References 41 publications

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“…In this new conformation, cyclohexanone is now located over the flavin in a position that enables formation of the Criegee intermediate. In combination with new structures (so-called 'tight' and 'loose' CHMO complexes [32]) reported for CHMO -product (e-caprolactone) complexes, these ligand complexes emphasize the dynamic nature of the CHMO active site and the conformational transitions that are required to facilitate the chemical changes throughout the complex reaction cycles of BVMOs.…”

Section: Catalytic Cycles and New Chemistriesmentioning

confidence: 89%

Sweating the assets of flavin cofactors: new insight of chemical versatility from knowledge of structure and mechanism

Leys

Scrutton

2016

Current Opinion in Structural Biology

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Flavins are arguably one of the most versatile cofactors by virtue of the reactivity of the isoalloxazine ring system. A varied catalogue of reactions for the diverse family of flavoenzymes has been reported, leading to unifying concepts in (long-range) electron transfer, oxygen activation, photochemistry and substrate redox reactions. Recent examples of unprecedented flavin chemistry have been reported that uncover hidden depths of the flavoenzyme chemical repertoire. These include ring expansion of flavin through prenylation and formation of the superoxidized flavin-N5 oxide species. These and other new flavin based species are reviewed here and suggest further exciting discoveries await the flavoenzymology field.

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Section: Catalytic Cycles and New Chemistriesmentioning

confidence: 89%

Sweating the assets of flavin cofactors: new insight of chemical versatility from knowledge of structure and mechanism

Leys

Scrutton

2016

Current Opinion in Structural Biology

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“…HI-31; GenBank BAH56677) as well as the results of directed evolution, 17 amino acid residues, illustrated in Figure S1 with hollow blue arrows, are close to the active site and contribute to the enantioselectivity. 19, 20, 54–56 …”

Section: Resultsmentioning

confidence: 99%

Substitution of a Single Amino Acid Reverses the Regiospecificity of the Baeyer–Villiger Monooxygenase PntE in the Biosynthesis of the Antibiotic Pentalenolactone

Chen

Zhu

et al. 2016

Biochemistry

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In the biosynthesis of pentalenolactone (1), PenE and PntE, orthologous proteins from Streptomyces exfoliatus and S. arenae, respectively, catalyze the flavin-dependent Baeyer-Villiger oxidation of 1-deoxy-11-oxopentalenic acid (4) to the lactone pentalenolactone D (5), in which the less-substituted methylene carbon has migrated. By contrast, the paralogous PtlE enzyme from S. avermitilis catalyzes the oxidation of 4 to neopentalenolactone D (6), in which the more substituted methane substitution has undergone migration. We report the design and analysis of 13 single and multiple mutants of PntE mutants in order to identify the key amino acids that contribute to the regiospecificity of these two classes of Baeyer-Villiger monooxygenases. The L185S mutation in PntE reversed the observed regiospecificity of PntE such that all recombinant PntE mutants harboring this L185S mutation acquired the characteristic regiospecificity of PtlE, catalyzing the conversion of 4 to 6 as the major product. The recombinant PntE mutant harboring R484L exhibited reduced regiospecificity, generating a mixture of lactones containing more than 17% of 6. These in vitro results were corroborated by analysis of the complementation of the S. avermitilis ΔptlED double deletion mutant with pntE mutants, such that pntE mutants harboring L185S produced 6 as the major product, while complemention of the ΔptlED deletion mutant with pntE mutants carrying the R484L mutation gave 6 as more than 33% of the total lactone product mixture.

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“…The BVMO Lepto consists of 488 amino acids and lacks 52 residues that comprise one helical turn and two helices in the polypeptide chain of the CHMO from Rhodococcus sp. HI-31 (Yachnin et al 2014) as predicted by homology modeling (Additional file 1: Figures S1, S2). This is a feature that BVMO Lepto shares with the others BVMOs of the same clade (Fig.…”

Section: Discussionmentioning

confidence: 99%

Cloning and characterization of the Type I Baeyer–Villiger monooxygenase from Leptospira biflexa

et al. 2017

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Baeyer–Villiger monooxygenases are recognized by their ability and high selectivity as oxidative biocatalysts for the generation of esters or lactones using ketones as starting materials. These enzymes represent valuable tools for biooxidative syntheses since they can catalyze reactions that otherwise involve strong oxidative reagents. In this work, we present a novel enzyme, the Type I Baeyer–Villiger monooxygenase from Leptospira biflexa. This protein is phylogenetically distant from other well-characterized BVMOs. In order to study this new enzyme, we cloned its gene, expressed it in Escherichia coli and characterized the substrate scope of the Baeyer–Villiger monooxygenase from L. biflexa as a whole-cell biocatalyst. For this purpose, we performed the screening of a collection of ketones with variable structures and sizes, namely acyclic ketones, aromatic ketones, cyclic ketones, and fused ketones. As a result, we observed that this biocatalyst readily oxidized linear- and branched- medium-chain ketones, alkyl levulinates and linear ketones with aromatic substituents with excellent regioselectivity. In addition, this enzyme catalyzed the oxidation of 2-substituted cycloketone derivatives but showed an unusual selection against substituents in positions 3 or 4 of the ring.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0390-5) contains supplementary material, which is available to authorized users.

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

Cited by 39 publications

References 41 publications

Sweating the assets of flavin cofactors: new insight of chemical versatility from knowledge of structure and mechanism

Sweating the assets of flavin cofactors: new insight of chemical versatility from knowledge of structure and mechanism

Substitution of a Single Amino Acid Reverses the Regiospecificity of the Baeyer–Villiger Monooxygenase PntE in the Biosynthesis of the Antibiotic Pentalenolactone

Cloning and characterization of the Type I Baeyer–Villiger monooxygenase from Leptospira biflexa

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