EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.14.21 | gene dszC, sequence comparisons, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) | Rhodococcus erythropolis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.14.14.21 | purified recombinant His-taged enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of 20 mg/ml protein in 20 mM Tris-HCl (pH 8.0) and 150 mM NaCl, with FMN in a 1:10 molar ratio, with 0.002 ml of reservoir solution containing 200 mM lithium sulfate, 100 mM Bis-Tris, pH 6.5, and 35% w/v PEG 3350, equilibration against 0.3 ml reservoir solution, 4°C, X-ray diffraction structure determination and analysis at 2.26 A resolution, molecular replacement using DszC structure, PDB ID 4JEK, as the search model | Rhodococcus erythropolis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.14.14.21 | F161A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | F250A | site-directed mutagenesis,almost inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | F250R | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | F415A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Rhodococcus erythropolis |
1.14.14.21 | H391A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | additional information | construction of a truncated mutant lacking the last eight residues of the C-terminus, the truncated mutant does not exhibit catalytic activity and primarily forms 50-kDa monomers in analytical ultracentrifugation analysis | Rhodococcus erythropolis |
1.14.14.21 | N129A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | R338A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | S163A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | S215A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | S417A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rhodococcus erythropolis |
1.14.14.21 | T416A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Rhodococcus erythropolis |
1.14.14.21 | W205A | site-directed mutagenesis, inactive mutant | Rhodococcus erythropolis |
1.14.14.21 | Y96A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rhodococcus erythropolis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.14.21 | dibenzothiophene + 2 FMNH2 + 2 O2 | Rhodococcus erythropolis | overall reaction | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
1.14.14.21 | dibenzothiophene + FMNH2 + O2 | Rhodococcus erythropolis | - |
dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
1.14.14.21 | dibenzothiophene-5-oxide + FMNH2 + O2 | Rhodococcus erythropolis | - |
dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.14.21 | Rhodococcus erythropolis | - |
gene dszC | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.14.14.21 | recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration | Rhodococcus erythropolis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.14.21 | dibenzothiophene + 2 FMNH2 + 2 O2 | overall reaction | Rhodococcus erythropolis | dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O | - |
? | |
1.14.14.21 | dibenzothiophene + FMNH2 + O2 | - |
Rhodococcus erythropolis | dibenzothiophene-5-oxide + FMN + H2O | - |
? | |
1.14.14.21 | dibenzothiophene-5-oxide + FMNH2 + O2 | - |
Rhodococcus erythropolis | dibenzothiophene-5,5-dioxide + FMN + H2O | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.14.21 | DBT monooxygenase | - |
Rhodococcus erythropolis |
1.14.14.21 | dszC | - |
Rhodococcus erythropolis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.14.21 | FMNH2 | - |
Rhodococcus erythropolis | |
1.14.14.21 | NADH | DszD is responsible for supplying reducing equivalents in the form of FMNH2 to the monooxygenase DszC | Rhodococcus erythropolis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.14.14.21 | metabolism | DBT monooxygenase from Rhodococcus erythropolis is involved in the first step of the 4S pathway. Dibenzothiophene and its derivatives are resistant to the hydrodesulfurization method often used in industry, but they are susceptible to enzymatic desulfurization via the 4S pathway | Rhodococcus erythropolis |
1.14.14.21 | additional information | the C-terminus (410-417) of enzyme DszC, which is located in the interior of the protein, is important for the stabilization of the active conformation of the substrate-binding pocket and the tetrameric state and plays a significant role in the catalytic activity of the enzyme. The residues around the site are conserved: Tyr96, Asn129, Phe161, Ser163, Trp205, Ser215, Phe250, and His391 | Rhodococcus erythropolis |