BRENDA - Enzyme Database

Isolation, cloning and characterization of a tyrosinase with improved activity in organic solvents from Bacillus megaterium

Shuster, V.; Fishman, A.; J. Mol. Microbiol. Biotechnol. 17, 188-200 (2009)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.14.18.1
2-propanol
activity is enhanced in the presence of 10-50% (v/v)
Bacillus megaterium
1.14.18.1
DMSO
activity is enhanced in the presence of 10-50% (v/v), the activity in 30% (v/v) DMSO is 170% of the activity in water and the enantioselectivity towards L-DOPA decreases by 40%
Bacillus megaterium
1.14.18.1
ethanol
activity is enhanced in the presence of 10-50% (v/v)
Bacillus megaterium
1.14.18.1
methanol
activity is enhanced in the presence of 10-50% (v/v), the residual activity following an incubation period of 17 h in 0-70% methanol is constant
Bacillus megaterium
Cloned(Commentary)
EC Number
Commentary
Organism
1.14.18.1
expressed in Escherichia coli BL21(DE3) cells
Bacillus megaterium
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.18.1
beta-mercaptoethanol
complete inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
EDTA
27% inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
glutathione
complete inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
Sodium diethyldithiocarbamate
complete inhibition at 1.0 mM
Bacillus megaterium
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.18.1
0.075
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
0.35
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
0.73
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
1.18
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
1.7
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.18.1
31000
-
gel filtration
Bacillus megaterium
1.14.18.1
35000
-
SDS-PAGE
Bacillus megaterium
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.18.1
Bacillus megaterium
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.18.1
Ni2+-bound affinity column chromatography, gel filtration
Bacillus megaterium
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.18.1
(+)-catechin + O2
60.8% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
(+/-)-catechin + O2
58.7% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
3-aminophenol + O2
3.9% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
caffeic acid + O2
60.9% activity compared to L-DOPA
699628
Bacillus megaterium
caffeoyl quinone + H2O
-
-
-
?
1.14.18.1
catechol + O2
10.3% activity compared to L-DOPA
699628
Bacillus megaterium
1,2-benzoquinone + H2O
-
-
-
?
1.14.18.1
chlorogenic acid + O2
20.1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
D-DOPA + O2
34.0% activity compared to L-DOPA
699628
Bacillus megaterium
D-dopaquinone + H2O
-
-
-
?
1.14.18.1
D-tyrosine + O2
20.3% activity compared to L-DOPA
699628
Bacillus megaterium
D-dopa + H2O
-
-
-
?
1.14.18.1
DL-DOPA + O2
78% activity compared to L-DOPA
699628
Bacillus megaterium
DL-dopaquinone + H2O
-
-
-
?
1.14.18.1
DL-tyrosine + O2
20.6% activity compared to L-DOPA
699628
Bacillus megaterium
DL-DOPA + H2O
-
-
-
?
1.14.18.1
epicatechin + O2
23.4% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
L-Dopa + O2
100% activity
699628
Bacillus megaterium
L-dopaquinone + H2O
-
-
-
?
1.14.18.1
L-tyrosine + O2
35.6% activity compared to L-DOPA
699628
Bacillus megaterium
L-DOPA + H2O
-
-
-
?
1.14.18.1
additional information
vanillic acid, 1-naphthol, 2,6-dimethoxyphenol, and resorcinol are no substrates for tyrosinase
699628
Bacillus megaterium
?
-
-
-
-
1.14.18.1
o-coumaric acid + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
p-coumaric acid + O2
6.1% activity compared to L-DOPA
699628
Bacillus megaterium
caffeic acid + H2O
-
-
-
?
1.14.18.1
p-hydroxybenzoic acid + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
phenol + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
phloroglucin + O2
1.6% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
pyrogallol + O2
99.9% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
tyramine + O2
2.5% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.18.1
monomer
1 * 31000, gel filtration
Bacillus megaterium
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.18.1
50
-
the maximum activity for both monophenolase and diphenolase activities of tyrosinase is observed at 50°C
Bacillus megaterium
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
1.14.18.1
10
70
-
Bacillus megaterium
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.14.18.1
50
70
the enzyme is fully stable below 50°C, but rapidly loses its activity at 70°C
Bacillus megaterium
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.18.1
2.5
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
6
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
10.1
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
12.12
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
21.3
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.18.1
7
-
the maximum activity for both monophenolase and diphenolase activities of tyrosinase is observed at pH 7.0
Bacillus megaterium
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.14.18.1
4
10
for monophenolase activity of tyrosinase
Bacillus megaterium
1.14.18.1
4
7
for diphenolase activity of tyrosinase
Bacillus megaterium
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.14.18.1
2-propanol
activity is enhanced in the presence of 10-50% (v/v)
Bacillus megaterium
1.14.18.1
DMSO
activity is enhanced in the presence of 10-50% (v/v), the activity in 30% (v/v) DMSO is 170% of the activity in water and the enantioselectivity towards L-DOPA decreases by 40%
Bacillus megaterium
1.14.18.1
ethanol
activity is enhanced in the presence of 10-50% (v/v)
Bacillus megaterium
1.14.18.1
methanol
activity is enhanced in the presence of 10-50% (v/v), the residual activity following an incubation period of 17 h in 0-70% methanol is constant
Bacillus megaterium
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.18.1
expressed in Escherichia coli BL21(DE3) cells
Bacillus megaterium
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.18.1
beta-mercaptoethanol
complete inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
EDTA
27% inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
glutathione
complete inhibition at 1.0 mM
Bacillus megaterium
1.14.18.1
Sodium diethyldithiocarbamate
complete inhibition at 1.0 mM
Bacillus megaterium
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.18.1
0.075
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
0.35
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
0.73
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
1.18
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
1.7
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.18.1
31000
-
gel filtration
Bacillus megaterium
1.14.18.1
35000
-
SDS-PAGE
Bacillus megaterium
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.18.1
Ni2+-bound affinity column chromatography, gel filtration
Bacillus megaterium
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.18.1
(+)-catechin + O2
60.8% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
(+/-)-catechin + O2
58.7% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
3-aminophenol + O2
3.9% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
caffeic acid + O2
60.9% activity compared to L-DOPA
699628
Bacillus megaterium
caffeoyl quinone + H2O
-
-
-
?
1.14.18.1
catechol + O2
10.3% activity compared to L-DOPA
699628
Bacillus megaterium
1,2-benzoquinone + H2O
-
-
-
?
1.14.18.1
chlorogenic acid + O2
20.1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
D-DOPA + O2
34.0% activity compared to L-DOPA
699628
Bacillus megaterium
D-dopaquinone + H2O
-
-
-
?
1.14.18.1
D-tyrosine + O2
20.3% activity compared to L-DOPA
699628
Bacillus megaterium
D-dopa + H2O
-
-
-
?
1.14.18.1
DL-DOPA + O2
78% activity compared to L-DOPA
699628
Bacillus megaterium
DL-dopaquinone + H2O
-
-
-
?
1.14.18.1
DL-tyrosine + O2
20.6% activity compared to L-DOPA
699628
Bacillus megaterium
DL-DOPA + H2O
-
-
-
?
1.14.18.1
epicatechin + O2
23.4% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
L-Dopa + O2
100% activity
699628
Bacillus megaterium
L-dopaquinone + H2O
-
-
-
?
1.14.18.1
L-tyrosine + O2
35.6% activity compared to L-DOPA
699628
Bacillus megaterium
L-DOPA + H2O
-
-
-
?
1.14.18.1
additional information
vanillic acid, 1-naphthol, 2,6-dimethoxyphenol, and resorcinol are no substrates for tyrosinase
699628
Bacillus megaterium
?
-
-
-
-
1.14.18.1
o-coumaric acid + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
p-coumaric acid + O2
6.1% activity compared to L-DOPA
699628
Bacillus megaterium
caffeic acid + H2O
-
-
-
?
1.14.18.1
p-hydroxybenzoic acid + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
phenol + O2
less than 1% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
phloroglucin + O2
1.6% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
pyrogallol + O2
99.9% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
1.14.18.1
tyramine + O2
2.5% activity compared to L-DOPA
699628
Bacillus megaterium
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.18.1
monomer
1 * 31000, gel filtration
Bacillus megaterium
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.18.1
50
-
the maximum activity for both monophenolase and diphenolase activities of tyrosinase is observed at 50°C
Bacillus megaterium
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
1.14.18.1
10
70
-
Bacillus megaterium
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.14.18.1
50
70
the enzyme is fully stable below 50°C, but rapidly loses its activity at 70°C
Bacillus megaterium
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.18.1
2.5
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
6
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
10.1
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
12.12
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
21.3
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.18.1
7
-
the maximum activity for both monophenolase and diphenolase activities of tyrosinase is observed at pH 7.0
Bacillus megaterium
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.14.18.1
4
10
for monophenolase activity of tyrosinase
Bacillus megaterium
1.14.18.1
4
7
for diphenolase activity of tyrosinase
Bacillus megaterium
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.18.1
5.1
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
7.1
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
28.9
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
29.2
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
32.9
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.18.1
5.1
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
7.1
-
D-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
28.9
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium
1.14.18.1
29.2
-
L-Dopa
in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO
Bacillus megaterium
1.14.18.1
32.9
-
L-tyrosine
in 50 mM potassium phosphate buffer (pH 7.0)
Bacillus megaterium