belongs to group of Bayer-Villiger monooxygenases of the NADH plus FMN-dependent type 2 enzymes; catalysis of 2 mechanistically different types of biochemical reactions within the confines of the same active site; cubic space active site model, topography; higher activity in oxidation of sulfides to the corresponding sulfoxides than in lactonization of ketones; isozyme, enantiocomplementary and isofunctional isozymes 2,5-DKCMO EC 1.14.15.2 and 3,6-DKCMO EC 1.14.15.x; mechanism
i.e. cyclopentaneacetic acid, 3-hydroxy-2,2,3-trimethyl-5-oxo, delta-lactone, reacts sponaneously to (2,2-dimethyl-5-oxo-cyclopent-3-en-1-yl)acetic acid
i.e. 3,6-DKCMO; isoform, which is probably a different enzyme; isozyme, enantiocomplementary and isofunctional isozymes 2,5-DKCMO EC 1.14.15.2 and 3,6-DKCMO EC 1.14.15.x
isoform, which is probably a different enzyme; isozyme, enantiocomplementary and isofunctional isozymes 2,5-DKCMO EC 1.14.15.2 and 3,6-DKCMO EC 1.14.15.x
the orientation of the isoalloxazine ring of the FMN cofactor in the active site of the TIM-barrel fold enzyme differs significantly from that previously observed in enzymes of the bacterial luciferase-like superfamily. The Ala77 residue is in a cis conformation and forms a beta-bulge at the C-terminus of beta-strand 3, which is a feature observed in many proteins of this superfamily. Both the 2,5-DKMO and 3,6-DKMO oxygenating components have sequence similarity to bacterial luciferases and bear little similarity to type I Baeyer-Villiger monooxygenase, type I BVMOs
type 2 Baeyer-Villiger monooxygenases (type 2 BVMOs) are a subgroup of the NAD(P)H:FMN-dependent two-component monooxygenases (TCMOs). They can alternatively be classed as a subgroup of the NAD(P)H:FMN-dependent class C flavoprotein monooxygenases
type 2 Baeyer-Villiger monooxygenases (type 2 BVMOs) are a subgroup of the NAD(P)H:FMN-dependent two-component monooxygenases (TCMOs). They can alternatively be classed as a subgroup of the NAD(P)H:FMN-dependent class C flavoprotein monooxygenases
2,5- and 3,6-diketocamphane monooxygenase (DKCMO) are two enantiocomplementary isoenzymes that catalyse a key lactone-forming step in the degradation of the (+)- and (-)-camphor antipodes, respectively, in Pseudomonas putida NCIMB 10007. Enzyme 3,6-diketocamphane monooxygenase distributes the flavin nucleotide- and nicotinamide nucleotide-dependent tasks between a homodimeric monooxygenase component and a separate flavin reductase (FR) with unbound FMN, the flavin thus effectively serving as a second substrate to transfer reducing power between the functionally distinct subunits. Various flavin reductases function effectively as sources of the requisite FMNH2 to 3,6-diketocamphane monooxygenase at different times throughout growth on camphor, significant subsequent contribution throughout the mid- to late-exponential phases of growth is also made by the camphor-induced homodimeric 37.0 kDa flavin reductase Fred, possible involvement of camphor-induced putidaredoxin reductase as a contributory activity. Analysis of flavin reductases in Pseudomonas putida NCIMB 10007, overview
2,5- and 3,6-diketocamphane monooxygenase (DKCMO) are two enantiocomplementary isoenzymes that catalyse a key lactone-forming step in the degradation of the (+)- and (-)-camphor antipodes, respectively, in Pseudomonas putida NCIMB 10007. Enzyme 3,6-diketocamphane monooxygenase distributes the flavin nucleotide- and nicotinamide nucleotide-dependent tasks between a homodimeric monooxygenase component and a separate flavin reductase (FR) with unbound FMN, the flavin thus effectively serving as a second substrate to transfer reducing power between the functionally distinct subunits. Various flavin reductases function effectively as sources of the requisite FMNH2 to 3,6-diketocamphane monooxygenase at different times throughout growth on camphor, significant subsequent contribution throughout the mid- to late-exponential phases of growth is also made by the camphor-induced homodimeric 37.0 kDa flavin reductase Fred, possible involvement of camphor-induced putidaredoxin reductase as a contributory activity. Analysis of flavin reductases in Pseudomonas putida NCIMB 10007, overview
i.e. cyclopentaneacetic acid, 3-hydroxy-2,2,3-trimethyl-5-oxo, delta-lactone, i.e. 3,4,4-trimethyl-6-carboxy-methyl-DELTA3-cyclopentenone, product is an unstable lactone-intermediate and forms spontaneously (2,2-dimethyl-5-oxo-cyclopent-3-en-1-yl)acetic acid, i.e. 2-oxo-DELTA3-4,5,5-trimethylcyclopentenyl acetic acid
catalyzes stereoselective electrophilic biooxidation of a wide range of prochiral organic sulfoxides to the corresponding chiral sulfoxides as well as the nucleophilic biooxidation of ketones to lactones with different enantio- and stereoselectivity, overview
catalyzes stereoselective electrophilic biooxidation of a wide range of prochiral organic sulfoxides to the corresponding chiral sulfoxides as well as the nucleophilic biooxidation of ketones to lactones with different enantio- and stereoselectivity, overview
dependent on, FMN cofactor modelling. Type II Baeyer-Villiger monooxygenases are attributed to the R group in relation to the re-face attachment of the hydroperoxide to the flavin coenzyme. The FMN cofactor binds in the cleft formed at the C-terminal side of the TIM barrel, with the phosphate group involved in hydrogen bonding at the edge of the cleft and the isoalloxazine ring located deep inside of the cleft
growth on (+)-, (-)- and (rac)-camphor. The two DKCMO isoenzymes, 2,5- and 3,6-diketocamphane monooxygenase, are highly selective towards their respective antipodes when serving as substrates for biooxygenation, nevertheless a small but significant cross-inducibility of the enantiomerically redundant DKCMO isoenzyme by both camphor antipodes exists
growth on (+)-, (-)- and (rac)-camphor. The two DKCMO isoenzymes, 2,5- and 3,6-diketocamphane monooxygenase, are highly selective towards their respective antipodes when serving as substrates for biooxygenation, nevertheless a small but significant cross-inducibility of the enantiomerically redundant DKCMO isoenzyme by both camphor antipodes exists
purified recombinant N-terminally His6-tagged enzyme, by Microbatch crystallization, mixing of 7 mg/ml protein in 20 mM FMN, 5 mM NADH and 5 mM (-)-camphor in a 1:1 ration, purified native enzyme, by vapour-diffusion technique, 10 mg/ml protein solution are mixed in equal volumes with 50 mM PIPES pH 6.5, 50% ammonium sulfate, room temperature, best from 100 mM HEPES pH 7.0, 20% PEG 3350 in the presence of 20 mM FMN, 5 mM NADH and 5 mM (-)-camphor, at 18°C, X-ray diffraction structure determination and analysis at 1.9-2.7 A resolution, the enzyme's crystal structure is solved by a combination of multiple anomalous dispersion from a bromine crystal soak and molecular replacement using a bacterial luciferase model
biooxygenating subunits of both enantiospecific DKCMO isoenzymes to homogeneity by a preparative process involving anion exchange chromatography and ultrafiltration, purification and detection of FMN-reductase activities
the enzyme is encoded on the large CAM plasmid of Pseudomonas putida, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Biotransformation of organic sulfides. Predictive active site models for sulfoxidation catalyzed by 2,5-diketocamphane 1,2-monooxygenase and 3,6-diketocamphane 1,6-monooxygenase, enantiocomplementary enzymes from Pseudomonas putida NCIMB 10007
The purification and crystallization of 2,5-diketocamphane 1,2-monooxygenase and 3,6-diketocamphane 1,6-monooxygenase from Pseudomonas putida NCIMB 10007
Kadow, M.; Loschinski, K.; Sass, S.; Schmidt, M.; Bornscheuer, U.
Completing the series of BVMOs involved in camphor metabolism of Pseudomonas putida NCIMB 10007 by identification of the two missing genes, their functional expression in E. coli, and biochemical characterization
The oxygenating constituent of 3,6-diketocamphane monooxygenase from the CAM plasmid of Pseudomonas putida the first crystal structure of a type II Baeyer-Villiger monooxygenase