show A novel enzyme, 2'-hydroxybiphenyl-2-sulfinate desulfinase (DszB), from a dibenzothiophene-desulfurizing bacterium Rhodococcus erythropolis KA2-5-1: gene overexpression and enzyme characterization
Nakayama, N.; Matsubara, T.; Ohshiro, T.; Moroto, Y.; Kawata, Y.; Koizumi, K.; Hirakawa, Y.; Suzuki, M.; Maruhashi, K.; Izumi, Y.; Kurane, R.; Biochim. Biophys. Acta 1598, 122-130 (2002)
Data extracted from this reference:
Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
3.13.1.3
additional information
no activation by metal ions
Rhodococcus erythropolis
Cloned(Commentary)
EC Number
Commentary
Organism
3.13.1.3
gene dszZ, DNA sequence determination, cooverexpression in Escherichia coli with chaperonin genes groEL and groES at 25°C
Rhodococcus erythropolis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.13.1.3
C26S
site-directed mutagenesis of the only cysteine residue results in an completely inactive mutant
Rhodococcus erythropolis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.13.1.3
2,2'-Dihydroxybiphenyl
competitive
Rhodococcus erythropolis
3.13.1.3
2,3-Dihydroxybiphenyl
competitive
Rhodococcus erythropolis
3.13.1.3
2-Hydroxybiphenyl
competitive, product inhibition
Rhodococcus erythropolis
3.13.1.3
Ag+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
Cu2+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
DTNB
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
Hg2+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
additional information
no inhibition by product sulfite, no inhibition by chelating agents EDTA, 1,10-phenanthroline, and 8-hydroxyquinone
Rhodococcus erythropolis
3.13.1.3
N-ethylmaleimide
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
p-chloromercuribenzoate
strong inhibition at 1 mM
Rhodococcus erythropolis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.13.1.3
0.0082
-
2'-hydroxybiphenyl-2-sulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
3.13.1.3
0.062
-
2-phenylbenzenesulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.13.1.3
additional information
no metal ions required
Rhodococcus erythropolis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.13.1.3
40000
-
1 * 40000, SDS-PAGE
Rhodococcus erythropolis
3.13.1.3
43000
-
gel filtration
Rhodococcus erythropolis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
Rhodococcus erythropolis
-
2-hydroxybiphenyl + sulfite
-
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
Rhodococcus erythropolis KA2-5-1
-
2-hydroxybiphenyl + sulfite
-
Rhodococcus erythropolis KA2-5-1
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.2.24
Rhodococcus erythropolis
-
-
-
3.1.2.24
Rhodococcus erythropolis KA2-5-1
-
-
-
3.13.1.3
Rhodococcus erythropolis
-
-
-
3.13.1.3
Rhodococcus erythropolis KA2-5-1
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.13.1.3
recombinant 7.8fold from Escherichia coli, and native from strain KA2-5-1 160fold, to homogeneity
Rhodococcus erythropolis
Reaction
EC Number
Reaction
Commentary
Organism
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O = 2-hydroxybiphenyl + sulfite
Cys26 is essential for activity
Rhodococcus erythropolis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.13.1.3
0.18
-
purified native enzyme
Rhodococcus erythropolis
3.13.1.3
0.207
-
purified recombinant enzyme
Rhodococcus erythropolis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
-
650387
Rhodococcus erythropolis
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
highly specific for
650387
Rhodococcus erythropolis
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
-
650387
Rhodococcus erythropolis KA2-5-1
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
highly specific for
650387
Rhodococcus erythropolis KA2-5-1
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
3.13.1.3
2-phenylbenzenesulfinate + H2O
-
650387
Rhodococcus erythropolis
biphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2-phenylbenzenesulfinate + H2O
-
650387
Rhodococcus erythropolis KA2-5-1
biphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
Subunits
EC Number
Subunits
Commentary
Organism
3.13.1.3
monomer
1 * 40000, SDS-PAGE
Rhodococcus erythropolis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.13.1.3
35
-
recombinant enzyme
Rhodococcus erythropolis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.13.1.3
28
-
recombinant enzyme, stable
Rhodococcus erythropolis
3.13.1.3
30
-
recombinant enzyme, rapid loss of activity above, inactivation after 4 h
Rhodococcus erythropolis
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.2.24
0.103
-
2-phenylbenzenesulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
3.1.2.24
0.123
-
2'-hydroxybiphenyl-2-sulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.13.1.3
7.5
-
about, recombinant enzyme
Rhodococcus erythropolis
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
3.13.1.3
additional information
no cofactor required
Rhodococcus erythropolis
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.13.1.3
0.25
-
2-Hydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
3.13.1.3
0.4
-
2,3-Dihydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
3.13.1.3
2
-
2,2'-Dihydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
3.13.1.3
additional information
no activation by metal ions
Rhodococcus erythropolis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.13.1.3
gene dszZ, DNA sequence determination, cooverexpression in Escherichia coli with chaperonin genes groEL and groES at 25°C
Rhodococcus erythropolis
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
3.13.1.3
additional information
no cofactor required
Rhodococcus erythropolis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.13.1.3
C26S
site-directed mutagenesis of the only cysteine residue results in an completely inactive mutant
Rhodococcus erythropolis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.13.1.3
2,2'-Dihydroxybiphenyl
competitive
Rhodococcus erythropolis
3.13.1.3
2,3-Dihydroxybiphenyl
competitive
Rhodococcus erythropolis
3.13.1.3
2-Hydroxybiphenyl
competitive, product inhibition
Rhodococcus erythropolis
3.13.1.3
Ag+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
Cu2+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
DTNB
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
Hg2+
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
additional information
no inhibition by product sulfite, no inhibition by chelating agents EDTA, 1,10-phenanthroline, and 8-hydroxyquinone
Rhodococcus erythropolis
3.13.1.3
N-ethylmaleimide
strong inhibition at 1 mM
Rhodococcus erythropolis
3.13.1.3
p-chloromercuribenzoate
strong inhibition at 1 mM
Rhodococcus erythropolis
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.13.1.3
0.25
-
2-Hydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
3.13.1.3
0.4
-
2,3-Dihydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
3.13.1.3
2
-
2,2'-Dihydroxybiphenyl
pH 7.0, 28°C, recombinant enzyme
Rhodococcus erythropolis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.13.1.3
0.0082
-
2'-hydroxybiphenyl-2-sulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
3.13.1.3
0.062
-
2-phenylbenzenesulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.13.1.3
additional information
no metal ions required
Rhodococcus erythropolis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.13.1.3
40000
-
1 * 40000, SDS-PAGE
Rhodococcus erythropolis
3.13.1.3
43000
-
gel filtration
Rhodococcus erythropolis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
Rhodococcus erythropolis
-
2-hydroxybiphenyl + sulfite
-
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
Rhodococcus erythropolis KA2-5-1
-
2-hydroxybiphenyl + sulfite
-
Rhodococcus erythropolis KA2-5-1
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.13.1.3
recombinant 7.8fold from Escherichia coli, and native from strain KA2-5-1 160fold, to homogeneity
Rhodococcus erythropolis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.13.1.3
0.18
-
purified native enzyme
Rhodococcus erythropolis
3.13.1.3
0.207
-
purified recombinant enzyme
Rhodococcus erythropolis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
-
650387
Rhodococcus erythropolis
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
highly specific for
650387
Rhodococcus erythropolis
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
-
650387
Rhodococcus erythropolis KA2-5-1
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
3.13.1.3
2'-hydroxybiphenyl-2-sulfinate + H2O
highly specific for
650387
Rhodococcus erythropolis KA2-5-1
2-hydroxybiphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
3.13.1.3
2-phenylbenzenesulfinate + H2O
-
650387
Rhodococcus erythropolis
biphenyl + sulfite
-
650387
Rhodococcus erythropolis
?
3.13.1.3
2-phenylbenzenesulfinate + H2O
-
650387
Rhodococcus erythropolis KA2-5-1
biphenyl + sulfite
-
650387
Rhodococcus erythropolis KA2-5-1
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.13.1.3
monomer
1 * 40000, SDS-PAGE
Rhodococcus erythropolis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.13.1.3
35
-
recombinant enzyme
Rhodococcus erythropolis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.13.1.3
28
-
recombinant enzyme, stable
Rhodococcus erythropolis
3.13.1.3
30
-
recombinant enzyme, rapid loss of activity above, inactivation after 4 h
Rhodococcus erythropolis
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.2.24
0.103
-
2-phenylbenzenesulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
3.1.2.24
0.123
-
2'-hydroxybiphenyl-2-sulfinate
pH 7.0, 28°C
Rhodococcus erythropolis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.13.1.3
7.5
-
about, recombinant enzyme
Rhodococcus erythropolis