Cloned (Comment) | Organism |
---|---|
gene dszC, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Rhodococcus sp. |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type, selenomethionine-labeled, and mutant enzymes, hanging drop vapor diffusion method, mixing of 15-20 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 100 mM NaCl with an equal volume of reservoir solution containing 0.2 M L2SO4, 0.1 M Bis-Tris, pH 6.5, and 23% w/v PEG 3350, at 20°C, X-ray diffraction structure determination and analysis at 1.79 A resolution, molecular replacement and modelling | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
D392N | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
F161A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
H388F | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
H391F | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
H92F | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
N129A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
R282A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
R370A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
S163A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
W205A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
Y96A | site-directed mutagenesis, inactive mutant | Rhodococcus sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dibenzothiophene + 2 FMNH2 + 2 O2 | Rhodococcus sp. | overall reaction | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
dibenzothiophene + 2 FMNH2 + 2 O2 | Rhodococcus sp. XP | overall reaction | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
dibenzothiophene + FMNH2 + O2 | Rhodococcus sp. | - |
dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
dibenzothiophene + FMNH2 + O2 | Rhodococcus sp. XP | - |
dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
dibenzothiophene-5-oxide + FMNH2 + O2 | Rhodococcus sp. | - |
dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? | |
dibenzothiophene-5-oxide + FMNH2 + O2 | Rhodococcus sp. XP | - |
dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | Q6WNP1 | gene dszC | - |
Rhodococcus sp. XP | Q6WNP1 | gene dszC | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type, selenomethionine-labeled, and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dibenzothiophene + 2 FMNH2 + 2 O2 | overall reaction | Rhodococcus sp. | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
dibenzothiophene + 2 FMNH2 + 2 O2 | overall reaction | Rhodococcus sp. XP | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
dibenzothiophene + FMNH2 + O2 | - |
Rhodococcus sp. | dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
dibenzothiophene + FMNH2 + O2 | - |
Rhodococcus sp. XP | dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
dibenzothiophene-5-oxide + FMNH2 + O2 | - |
Rhodococcus sp. | dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? | |
dibenzothiophene-5-oxide + FMNH2 + O2 | - |
Rhodococcus sp. XP | dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? | |
additional information | reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of the C4a-(hydro)xyflavin, substrate binding and mechanism, molecular docking | Rhodococcus sp. | ? | - |
? | |
additional information | reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of the C4a-(hydro)xyflavin, substrate binding and mechanism, molecular docking | Rhodococcus sp. XP | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | enzyme DszC is a tightly associated homotetramer, which can be aptly described as a dimer of dimers | Rhodococcus sp. |
More | two distinct conformations occur in the flexible lid loops adjacent to the active site (residue 280-295, between helix alpha9 and alpha10), that are named open and closed state respectively, and might show the status of the free and ligand-bound DszC | Rhodococcus sp. |
Synonyms | Comment | Organism |
---|---|---|
DBT monooxygenase | - |
Rhodococcus sp. |
dibenzothiophene monooxygenase | - |
Rhodococcus sp. |
dszC | - |
Rhodococcus sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMNH2 | reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of C4a-(hydro)peroxyflavin, molecular docking simulation and modeling, overview | Rhodococcus sp. |
General Information | Comment | Organism |
---|---|---|
evolution | DszC with specificity for FMN makes a unique member of the flavin monooxygenase family | Rhodococcus sp. |
malfunction | site-directed mutagenesis study shows that mutations in the residues involved either in catalysis or in flavin or substrate-binding result in a complete loss of enzyme activity, suggesting that the accurate positions of flavin and substrate are crucial for the enzyme activity | Rhodococcus sp. |
additional information | two distinct conformations occur in the flexible lid loops adjacent to the active site (residue 280-295, between helix alpha9 and alpha10), that are named open and closed state, respectively, and might show the status of the free and ligand-bound DszC | Rhodococcus sp. |
physiological function | the dibenzothiophene (DBT) monooxygenase DszC, which is the key initiating enzyme in 4S metabolic pathway, catalyzes sequential sulphoxidation reaction of DBT to DBT sulfoxide (DBTO), then DBT sulfone (DBTO2). Residues H391, and D392 directly participate in catalysis. Residues H92, Y96, N129, F161, S163, W205, R282, R370, and H388 are involved in flavin or substrate-binding | Rhodococcus sp. |