Information on EC 1.14.14.21 - dibenzothiophene monooxygenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.14.21
-
RECOMMENDED NAME
GeneOntology No.
dibenzothiophene monooxygenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dibenzothiophene + 2 FMNH2 + 2 O2 = dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
overall reaction
-
-
-
dibenzothiophene + FMNH2 + O2 = dibenzothiophene-5-oxide + FMN + H2O
show the reaction diagram
(1a)
-
-
-
dibenzothiophene-5-oxide + FMNH2 + O2 = dibenzothiophene-5,5-dioxide + FMN + H2O
show the reaction diagram
(1b)
-
-
-
SYSTEMATIC NAME
IUBMB Comments
dibenzothiophene,FMNH2:oxygen oxidoreductase
This bacterial enzyme catalyses the first two steps in the desulfurization pathway of dibenzothiophenes, the oxidation of dibenzothiophene into dibenzothiophene sulfone via dibenzothiophene-5-oxide. The enzyme forms a two-component system with a dedicated NADH-dependent FMN reductase (EC 1.5.1.42) encoded by the dszD gene, which also interacts with EC 1.14.14.22, dibenzothiophene sulfone monooxygenase.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Gordonia sp. F.5.25.8
-
-
-
Manually annotated by BRENDA team
gene dszC
UniProt
Manually annotated by BRENDA team
gene dszC
UniProt
Manually annotated by BRENDA team
gene dszC
-
-
Manually annotated by BRENDA team
gene dszC
-
-
Manually annotated by BRENDA team
i.e. D-1, gene dszC
UniProt
Manually annotated by BRENDA team
strain IGTS8, differs only in two positions from strain KA2-5-1 sequence; gene dszC
UniProt
Manually annotated by BRENDA team
gene dszC
UniProt
Manually annotated by BRENDA team
gene sox/dszC
UniProt
Manually annotated by BRENDA team
gene dszC
UniProt
Manually annotated by BRENDA team
gene dszC
UniProt
Manually annotated by BRENDA team
isolate SA21, gene dszC
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-methyldibenzothiophene + 2 FMNH2 + 2 O2
1-methyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
-
-
-
-
?
2,7-diethylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
2,8-dimethyldibenzothiophene + 2 FMNH2 + 2 O2
2,8-dimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
2-ethylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
2-ethyldibenzothiophene + 2 FMNH2 + 2 O2
2-ethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
2-methylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
2-methyldibenzothiophene + 2 FMNH2 + 2 O2
2-methyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
3,4,6,7-tetramethyldibenzothiophene + 2 FMNH2 + 2 O2
3,4,6,7-tetramethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
3,4,6-trimethyldibenzothiophene + 2 FMNH2 + 2 O2
3,4,6-trimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
-
-
-
?
3,4-benzo-dibenzothiophene + 2 FMNH2 + 2 O2
3,4-benzo-dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
3-ethyldibenzothiophene + 2 FMNH2 + 2 O2
3-ethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
-
-
-
?
3-methylbenzothiophene + 2 FMNH2 + 2 O2
3-methylbenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
-
-
-
?
3-methylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
-
less than 10% of the activity with dibenzothiophene
-
-
?
3-methyldibenzothiophene + 2 FMNH2 + 2 O2
3-methyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
4,6-diethyldibenzothiophene + 2 FMNH2 + 2 O2
4,6-diethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
4,6-dimethyldibenzothiophene + 2 FMNH2 + 2 O2
4,6-dimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
4-methyldibenzothiophene + 2 FMNH2 + 2 O2
4-methyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
5-methylbenzothiophene + 2 FMNH2 + 2 O2
5-methylbenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
-
-
-
?
5-methylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
-
33% of the activity with dibenzothiophene
-
-
?
7-ethylbenzothiophene + FMNH2 + O2
?
show the reaction diagram
-
less than 10% of the activity with dibenzothiophene
-
-
?
benzothiophene + 2 FMNH2 + 2 O2
benzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
dibenzothiophene + 2 FADH2 + 2 O2
dibenzothiophene-5,5-dioxide + 2 FAD + 2 H2O
show the reaction diagram
dibenzothiophene + 2 FMNH2 + 2 O2
dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
dibenzothiophene + 2 reduced riboflavin + 2 O2
dibenzothiophene-5,5-dioxide + 2 riboflavin + 2 H2O
show the reaction diagram
dibenzothiophene + FMNH2 + O2
dibenzothiophene-5-oxide + FMN + H2O
show the reaction diagram
dibenzothiophene-5,5-dioxide + 2 FMNH2 + O2
2'-hydroxybiphenyl-2-sulfinate + 2 FMN + H2O
show the reaction diagram
dibenzothiophene-5-oxide + FMNH2 + O2
dibenzothiophene-5,5-dioxide + FMN + H2O
show the reaction diagram
thioxanthen-9-one + 2 FMNH2 + 2 O2
?
show the reaction diagram
thioxanthen-9-one + FMNH2 + O2
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2,8-dimethyldibenzothiophene + 2 FMNH2 + 2 O2
2,8-dimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
2-ethyldibenzothiophene + 2 FMNH2 + 2 O2
2-ethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
3,4,6,7-tetramethyldibenzothiophene + 2 FMNH2 + 2 O2
3,4,6,7-tetramethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
3,4,6-trimethyldibenzothiophene + 2 FMNH2 + 2 O2
3,4,6-trimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
A0SWL2
-
-
-
?
3-ethyldibenzothiophene + 2 FMNH2 + 2 O2
3-ethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
A0SWL2
-
-
-
?
4,6-dimethyldibenzothiophene + 2 FMNH2 + 2 O2
4,6-dimethyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
A0SWL2
-
-
-
?
4-methyldibenzothiophene + 2 FMNH2 + 2 O2
4-methyldibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
A0SWL2
-
-
-
?
benzothiophene + 2 FMNH2 + 2 O2
benzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
A0SWL2
-
-
-
?
dibenzothiophene + 2 FMNH2 + 2 O2
dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O
show the reaction diagram
dibenzothiophene + FMNH2 + O2
dibenzothiophene-5-oxide + FMN + H2O
show the reaction diagram
dibenzothiophene-5,5-dioxide + 2 FMNH2 + O2
2'-hydroxybiphenyl-2-sulfinate + 2 FMN + H2O
show the reaction diagram
dibenzothiophene-5-oxide + FMNH2 + O2
dibenzothiophene-5,5-dioxide + FMN + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
can substitute for NADH to some extent
riboflavin
low activity
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
2,2'-dithiobis(5-nitropyridine)
-
-
-
4-chloromercuribenzoate
-
-
4-chloromercuribenzoic acid
-
-
5,5'-dithio-bis-2-nitrobenzoic acid
94% inhibition at 0.5 mM
8-Quinolinol
-
-
CaCl2
16% inhibition at 1 mM
CoCl2
74% inhibition at 1 mM
CuCl2
complete inhibition at 1 mM
diethyldithiocarbamate
-
-
EDTA
68% inhibition at 1 mM
FAD
the flavin at high concentrations inhibits the activity of DszC due to autocatalytic oxidation of reduced flavin, formation of H2O2
FMN
the flavin at high concentrations inhibits the activity of DszC due to autocatalytic oxidation of reduced flavin, formation of H2O2
iodoacetate
46% inhibition at 1 mM
MnCl2
66% inhibition at 1 mM
N-ethylmaleimide
98% inhibition at 1 mM
p-chloromercuribenzoic acid
complete inhibition at 0.5 mM
ZnCl2
complete inhibition at 1 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FAD
stimulates enzyme DszC activity significantly at 0.01 mM, best at 0.035 mM
FMN
stimulates enzyme DszC activity significantly at 0.001 mM, best at 0.005 mM
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.36
benzothiophene
pH 7.0, 55°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 8
purified recombinant enzyme, pH 7.0, 30°C
18.6
purified enzyme, pH 7.0, 55°C
84.2
-
purified native enzyme, pH 7.0, 50°C
133
-
purified enzyme, pH 7.0, 30°C
163.2
-
purified recombinant enzyme, pH 7.0, 50°C
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 35
-
assay at
37 - 50
assay at, cell extract
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 65
activity range
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43000
4 * 43000, SDS-PAGE
44977
-
4 * 44977, sequence calculation
45800
x * 45800, about, sequnence calculation
86000
recombinant enzyme, gel filtration
180000
-
gel filtration
200000
250000
-
gel filtration
294000
recombinant enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homohexamer
homotetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme DszC, hanging drop vapour diffusion method, using a reservoir suliton containing 30% polyethylene glycol, 0.1 M sodium citrate, pH 5.6, and 0.2 M ammonium acetate, 4°C, X-ray diffraction structure determination and analysis
-
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
-
purified recombinant His-tagged enzyme in apoform and as FMN-bound enzyme DszC, two distinct conformations occur in the loop region (residues 131-142) adjacent to the active site, sitting drop vapor diffusion method, mixing of 0.001 ml protein solution with 0.001 ml of reservoir solution consisting of 0.2 M malonate, pH 6.0, 24% w/v PEG 3350, and 50 mM NaF, at 20°C, with or without 1 mM FMN, X-ray diffraction structure determination and analysis at 2.11 A an 2.3 A resolution, respectively, Each crystal contains two tetramers in the asymmetric unit, formed by two homodimers
-
purified recombinant wild-type and selenomethionine-labeled enzymes, hanging-drop vapour-diffusion method, mixing of 10 mg/ml wild-type enzyme in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl with reservoir solution containing 17.5% PEG 3350, 200 mM Bis-Tris, pH 7.5, and 200 mM ammonium sulfate, and mixing of 5 mg/ml selenomethionine-labeled enzyme in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl with reservoir solution containing 25% PEG 3350, 100 mM PIPES, pH 7.0, 200 mM ammonium sulfate, 20°C, X-ray diffraction structure determination and analysis at 2.4-2.9 A resolution
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
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 11
-
purified enzyme, stable at
736522
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
purified enzyme, 30 min, 80% activity remaning
65
purified enzyme, retains more than 80% of its activity after exposure for 30 min
70
purified enzyme, no activity remaining after 30 min
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 116fold from strain All-2 by two different steps of anion exchange chromatography and dialysis, followed by hydrophobic interaction and hydroxyapatite chromatography
native enzyme 19.2fold from strain WU-S2B by two different steps of anion exchange chromatography and one step of hydrophobic interaction chromatography, recombinant enzyme from Escherichia coli 4.5fold to homogeneity by anion exchange and hydrophobic interaction chromatography
-
native enzyme 24fold to homogeneity from strain D-1 by ammonium sulfate fractionation, anion exchange chromatography, hydrophobic interaction chromatography, and another different step of anion exchange chromatography, followed by gel filtration
-
native enzyme from strain IGTS8 by dye ligand affinity chromatography, dialysis, and hydrophobic interaction chromatography, followed by gel filtration
-
native enzyme to homogeneity by hydrophobic interaction and anion exchange chromatography. and gel filtration
-
recombinant enzyme 39fold from Escherichia coli strain BL21(DE3) by affinity chromatography
recombinant enzyme from Escherichia coli strain BL21 by anion exchange and hydrophobic interaction chromatography, followed by dialysis
recombinant enzyme from Escherichia coli strain BL21(DE3)
recombinant His-tagged enzyme from Escherichia coli strain DH5alpha by nickel affinty chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain Rosetta(DE3) by nickel affinity and anion exchange chromatography, followed by ultrafiltration
-
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration
-
recombinant N-terminally His-tagged enzyms from Escherichia coli strain by nickel affinity and anion exchange chromatography, and gel filtration
recombinant wild-type, selenomethionine-labeled, and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene bdsC, DNA and amino acid sequence determination and analysis, recombinant functional expression in Escherichia coli strain JM109, coexpression with a flavin reductase Frb, encoded by gene frb, UniProt ID Q75V96. Coexpression of frb with the DBT-desulfurization genes (bdsABC) from Bacillus subtilis strain WU-S2B is critical for high DBT-desulfurizing ability over a wide temperature range of 20-55°C, coexpression screening using indigo production as selective indication
gene bdsC, recombinant expression in Escherichia coli strain BL21(DE3)/pET21-a/bdsC
-
gene bdsC, recombinant overexpression in Escherichia coli
-
gene dszC
gene dszC, cloning from dszABC operon, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis
gene dszC, cloning from strain DS-3, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) and of selenomethionine-labeled enzyme in Escherichia coli strain B834(DE3)
gene dszC, DNA and amino acid sequence determination and analysis, cloning and recombinant expression in Escherichia coli strain JM109
gene dszC, DNA and amino acid sequence determination and analysis, sequence comparison, recombinant expression of His-tagged enzyme in Escherichia coli strain DH5alpha
-
gene dszC, functional recombinant enzyme expression in Thermus thermophilus strain HB27
-
gene dszC, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene dszC, phylogenetic analysis, recombinant expression in Escherichia coli strain BL21(DE3)
gene dszC, recombinant expression in Escherichia coli strain BL21(DE3)
gene dszC, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta(DE3)
-
gene dszC, recombinant expression of the complete dszABC operon, revealing that the ratio of mRNA quantity of dszA, dszB, and dszC is 11:3.3:1, the expression level of dszB is far lower than that of dszC. The termination codon of dszA and the initiation codon of dszB overlap, whereas there is a 13-bp gap between genes dszB and dszC. Quantitative analysis of dsz operon transcription by real-time PCR
gene dszC, sequence comparisons, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
gene sox/dszC, overexpression in Escherichia coli strain BL21
gene tdsC, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis, recombinant expression in Escherichia coli, coexpression with genes tdsC and tdsB, leading to established thermophilic desulfurization of dibenzothiophene in Escherichia coli host, an Escherichia coli oxidoreductase can be functionally coupled with the monooxygenases of a gram-positive thermophile
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F161A
-
site-directed mutagenesis, inactive mutant
F250A
-
site-directed mutagenesis,almost inactive mutant
F250R
-
site-directed mutagenesis, inactive mutant
F415A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
H391A
-
site-directed mutagenesis, inactive mutant
N129A
-
site-directed mutagenesis, inactive mutant
R338A
-
site-directed mutagenesis, inactive mutant
S163A
-
site-directed mutagenesis, inactive mutant
S215A
-
site-directed mutagenesis, inactive mutant
S417A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T416A
-
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
W205A
-
site-directed mutagenesis, inactive mutant
Y96A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D392N
site-directed mutagenesis, inactive mutant
F161A
site-directed mutagenesis, inactive mutant
H388F
site-directed mutagenesis, inactive mutant
H391F
site-directed mutagenesis, inactive mutant
H92F
site-directed mutagenesis, inactive mutant
N129A
site-directed mutagenesis, inactive mutant
R282A
site-directed mutagenesis, inactive mutant
R370A
site-directed mutagenesis, inactive mutant
S163A
site-directed mutagenesis, inactive mutant
W205A
site-directed mutagenesis, inactive mutant
Y96A
site-directed mutagenesis, inactive mutant
H92F
-
site-directed mutagenesis, inactive mutant
-
N129A
-
site-directed mutagenesis, inactive mutant
-
R282A
-
site-directed mutagenesis, inactive mutant
-
S163A
-
site-directed mutagenesis, inactive mutant
-
Y96A
-
site-directed mutagenesis, inactive mutant
-
additional information