The oxygenating constituent of 3,6-diketocamphane monooxygenase from the CAM plasmid of<i>Pseudomonas putida</i>: the first crystal structure of a type II Baeyer–Villiger monooxygenase

Isupov, Michail N.; Schröder, Ewald; Gibson, Ray; Beecher, Jean; Donadio, Giuliana; Saneei, Vahid; D'Cunha, Stephlina A.; McGhie, Emma J.; Sayer, C.; Davenport, Colin; Lau, Peter C. K.; Hasegawa, Yoshie; Iwaki, Hiroaki; Kadow, Maria; Balke, Kathleen; Bornscheuer, Uwe T.; Bourenkov, Gleb; Littlechild, Jennifer A.

doi:10.1107/s1399004715017939

Cited by 22 publications

(31 citation statements)

References 61 publications

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“…These data are particularly relevant to the recently proposed three-dimensional crystallographic structural model of 3,6-DKCMO from P. putida NCIMB 10007 [55]. The model predicts that there is no significant difference in the hydrogen bonding patterns of FMN and FNR in the active site of the enzyme, which does not correspond with the >500-fold difference in the actual K d values for the two forms of the flavin recorded with this enzyme.…”

Section: Discussionmentioning

confidence: 79%

Flavin-Dependent Redox Transfers by the Two-Component Diketocamphane Monooxygenases of Camphor-Grown Pseudomonas putida NCIMB 10007

Willetts

Kelly

2016

Microorganisms

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The progressive titres of key monooxygenases and their requisite native donors of reducing power were used to assess the relative contribution of various camphor plasmid (CAM plasmid)- and chromosome-coded activities to biodegradation of (rac)-camphor at successive stages throughout growth of Pseudomonas putida NCIMB 10007 on the bicylic monoterpenoid. A number of different flavin reductases (FRs) have the potential to supply reduced flavin mononucleotide to both 2,5- and 3,6-diketocamphane monooxygenase, the key isoenzymic two-component monooxygenases that delineate respectively the (+)- and (−)-camphor branches of the convergent degradation pathway. Two different constitutive chromosome-coded ferric reductases able to act as FRs can serve such as role throughout all stages of camphor-dependent growth, whereas Fred, a chromosome-coded inducible FR can only play a potentially significant role in the relatively late stages. Putidaredoxin reductase, an inducible CAM plasmid-coded flavoprotein that serves an established role as a redox intermediate for plasmid-coded cytochrome P450 monooxygenase also has the potential to serve as an important FR for both diketocamphane monooxygenases (DKCMOs) throughout most stages of camphor-dependent growth.

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Section: Discussionmentioning

confidence: 79%

Flavin-Dependent Redox Transfers by the Two-Component Diketocamphane Monooxygenases of Camphor-Grown Pseudomonas putida NCIMB 10007

Willetts

Kelly

2016

Microorganisms

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“…2WGK is a Type II Baeyer-Villiger monooxygenase from Pseudomonas putida [ 19 ], which has sequence coverage of ~ 98% and sequence identity of ~ 36%, with M . marinum MelF protein.…”

Section: Resultsmentioning

confidence: 99%

“…Sequence of MelF (Accession Number: gi|54289551.1) was PSI-BLASTed against PDB, and 2WGK with 36% sequence identity and 98% sequence coverage was found [ 19 ]. Yet Another Scientific Artificial Reality Application (YASARA) Dynamics and Structure (YASARA Biosciences GmBH, Vienna, Austria) (v 10.12.1), software was used for homology modelling and structural refinement ( Fig 1A ).…”

Section: Methodsmentioning

confidence: 99%

Rational design of drug-like compounds targeting Mycobacterium marinum MelF protein

et al. 2017

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The mycobacterial mel2 locus (mycobacterial enhanced infection locus, Rv1936-1941) is Mycobacterium marinum and M. tuberculosis specific, which can withstand reactive oxygen species (ROS) and reactive nitrogen species (RNS) induced stress. A library of over a million compounds was screened using in silico virtual ligand screening (VLS) to identify inhibitors against the modeled structure of MelF protein expressed by melF of mel2 locus so that M. marinum’s ability to withstand ROS/RNS stress could be reduced. The top ranked 1000 compounds were further screened to identify 178 compounds to maximize the scaffold diversity by manually evaluating the interaction of each compound with the target site. M. marinum melF was cloned, expressed and purified as maltose binding protein (MBP)-tagged recombinant protein in Escherichia coli. After establishing the flavin dependent oxidoreductase activity of MelF (~ 84 kDa), the inhibitors were screened for the inhibition of enzyme activity of whole cell lysate (WCL) and the purified MelF. Amongst these, 16 compounds could significantly inhibit the enzyme activity of purified MelF. For the six best inhibitory compounds, the minimal inhibitory concentration (MIC) was determined to be 3.4–19.4 μM and 13.5–38.8 μM for M. marinum and M. tuberculosis, respectively. Similarly, the minimal bactericidal concentration (MBC) was determined to be 6.8–38.8 μM and 27–38.8 μM against M. marinum and M. tuberculosis, respectively. One compound each in combination with isoniazid (INH) also showed synergistic inhibitory effect against M. marinum and M. tuberculosis with no cytotoxicity in HeLa cells. Interestingly, these inhibitors did not display any non-specific protein-structure destabilizing effect. Such inhibitors targeting the anti-ROS/RNS machinery may facilitate the efficient killing of replicating and nonreplicating mycobacteria inside the host cells.

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“…Type I monooxygenases are FAD and NADPH dependent biocatalysts and belong to the class B flavoenzymes, whereas Type II BVMOs require FMN and NADH for catalysis (Pazmino et al 2010b). Recently, the first crystal structure of a t y p e I I B V M O , n a m e l y 3 , 6 -d i k e t o c a m p h a n e monooxygenase, was published (Isupov et al 2015) which complements the intensive research in this area in the past years (Kadow et al , 2014. Nevertheless, most widely applied in synthetic chemistry are Type I BVMOs and more than 50 protein sequences are available for recombinant expression today.…”

Section: Baeyer-villiger Monooxygenases Classification Of Bvmosmentioning

confidence: 99%

Baeyer-Villiger oxidations: biotechnological approach

Bučko

Gemeiner

Schenkmayerová

et al. 2016

Appl Microbiol Biotechnol

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Baeyer-Villiger monooxygenases (BVMOs) are a very well-known and intensively studied class of flavin-dependent enzymes. Their substrate promiscuity, high chemo-, regio-, and enantioselectivity are prerequisites for the use in synthetic chemistry and should pave the way for successful industrial processes. Nonetheless, only a very limited number of industrial relevant transformations are known, mainly due to the lack of BVMOs stability and cofactor dependency. In this review, we focus on novel BVMO-mediated transformations, BVMOs in cascade type reactions, potential industrial applications, and how limitations have been tackled by the community. Special attention will be put on whole-cell immobilization strategies. We emphasize to bridge recent developments in fundamental research to industrial applications.

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The oxygenating constituent of 3,6-diketocamphane monooxygenase from the CAM plasmid ofPseudomonas putida: the first crystal structure of a type II Baeyer–Villiger monooxygenase

Cited by 22 publications

References 61 publications

Flavin-Dependent Redox Transfers by the Two-Component Diketocamphane Monooxygenases of Camphor-Grown Pseudomonas putida NCIMB 10007

Flavin-Dependent Redox Transfers by the Two-Component Diketocamphane Monooxygenases of Camphor-Grown Pseudomonas putida NCIMB 10007

Rational design of drug-like compounds targeting Mycobacterium marinum MelF protein

Baeyer-Villiger oxidations: biotechnological approach

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