BRENDA - Enzyme Database

In vitro activation of NAD-dependent alcohol dehydrogenases by Nudix hydrolases is more widespread than assumed

Ochsner, A.M.; Mueller, J.E.; Mora, C.A.; Vorholt, J.A.; FEBS Lett. 588, 2993-2999 (2014)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.244
Act
activator protein, UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase, in a pH-dependent manner. Act increases the Vmax of the enzyme and decreases the Km for methanol, the increase was 23fold at pH 7.4 and 50fold at pH 9.0
Bacillus methanolicus
1.1.1.244
activator protein ACT
UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase, in a pH-dependent manner. Act increases the Vmax of the enzyme and decreases the Km for methanol, the increase is 20fold at pH 7.4 and 9fold at pH 9.0
Bacillus methanolicus
1.1.1.244
additional information
the enzyme is activated by several Nudix hydrolases, relative activities of Adh enzymes, overview; the enzyme is activated by several Nudix hydrolases, relative activities of Adh enzymes, overview
Bacillus methanolicus
1.1.1.244
NudBC
a Nudix hydrolase from Bacillus coagulans, UniProt ID G2TKZ3; a Nudix hydrolase from Bacillus coagulans, UniProt ID G2TKZ3
Bacillus methanolicus
1.1.1.244
NudDK
a Nudix hydrolase from Desulfotomaculum kuznetsovii, UniProt ID F6CJI1; a Nudix hydrolase from Desulfotomaculum kuznetsovii, UniProt ID F6CJI1
Bacillus methanolicus
1.1.1.244
NudLF
a Nudix hydrolase from Lysinibacillus fusiformis, UniProt ID D7WXY3; a Nudix hydrolase from Lysinibacillus fusiformis, UniProt ID D7WXY3
Bacillus methanolicus
1.1.1.244
NudLS
a Nudix hydrolase from Lysinibacillus sphaericus, UniProt ID B1HRL7; a Nudix hydrolase from Lysinibacillus sphaericus, UniProt ID B1HRL7
Bacillus methanolicus
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
1.1.1.244
gene mdh2, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3); gene mdh, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Bacillus methanolicus
Engineering
EC Number
Protein Variants
Commentary
Organism
1.1.1.244
D38G
site-directed mutagenesis, the mutant is active with NADP+ and NAD+ in contrast to the wild-type enzyme
Bacillus methanolicus
1.1.1.244
D41G
site-directed mutagenesis, the mutant is active with NADP+ and NAD+ in contrast to the wild-type enzyme, it shows increased activity compared to the wild-type enzyme
Bacillus methanolicus
1.1.1.244
S101G
site-directed mutagenesis, the mutant shows altered kinetics, reduced activity, and altered pH-dependency compared to the wild-type enzyme, overview
Bacillus methanolicus
1.1.1.244
S98G
site-directed mutagenesis, the mutant shows altered kinetics, reduced activity, and altered pH-dependency compared to the wild-type enzyme, overview
Bacillus methanolicus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.244
additional information
-
additional information
Michaelis-Menten kinetics; Michaelis-Menten kinetics
Bacillus methanolicus
1.1.1.244
0.009
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.025
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.051
-
ethanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.096
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.137
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in presence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.15
-
methanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.161
-
ethanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.224
-
ethanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.225
-
ethanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.255
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.349
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.379
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in absence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.416
-
methanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.733
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.847
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
1.151
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1.1.1.244
methanol + NAD+
Bacillus methanolicus
-
formaldehyde + NADH + H+
-
-
r
1.1.1.244
methanol + NAD+
Bacillus methanolicus MGA3
-
formaldehyde + NADH + H+
-
-
r
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.1.1.244
Bacillus methanolicus
I3DTM5
-
-
1.1.1.244
Bacillus methanolicus
I3E2P9
-
-
1.1.1.244
Bacillus methanolicus MGA3
I3DTM5
-
-
1.1.1.244
Bacillus methanolicus MGA3
I3E2P9
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
1.1.1.244
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Bacillus methanolicus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.1.1.244
ethanol + NAD+
-
742514
Bacillus methanolicus
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.244
ethanol + NAD+
-
742514
Bacillus methanolicus MGA3
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NAD+
-
742514
Bacillus methanolicus
formaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NAD+
-
742514
Bacillus methanolicus MGA3
formaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus MGA3
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus MGA3
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
additional information
poor activity with 2-propanol and 1,2-popanediol
742514
Bacillus methanolicus
?
-
-
-
-
1.1.1.244
additional information
poor activity with 2-propanol and 1,2-popanediol
742514
Bacillus methanolicus MGA3
?
-
-
-
-
1.1.1.244
n-butanol + NAD+
-
742514
Bacillus methanolicus
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.244
n-butanol + NAD+
-
742514
Bacillus methanolicus MGA3
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.244
n-propanol + NAD+
-
742514
Bacillus methanolicus
propionaldehyde + NADH + H+
-
-
-
r
Synonyms
EC Number
Synonyms
Commentary
Organism
1.1.1.244
MDH
-
Bacillus methanolicus
1.1.1.244
MDH2
-
Bacillus methanolicus
1.1.1.244
NAD-dependent methanol dehydrogenase
-
Bacillus methanolicus
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.1.244
30
-
assay at; assay at
Bacillus methanolicus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.244
9
-
;
Bacillus methanolicus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.244
NAD+
;
Bacillus methanolicus
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.244
Act
activator protein, UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase, in a pH-dependent manner. Act increases the Vmax of the enzyme and decreases the Km for methanol, the increase was 23fold at pH 7.4 and 50fold at pH 9.0
Bacillus methanolicus
1.1.1.244
activator protein ACT
UniProt ID I3EA59, the in vitro activity of Bacillus methanolicus Mdh is increased by the endogenous activator protein Act, a Nudix hydrolase, in a pH-dependent manner. Act increases the Vmax of the enzyme and decreases the Km for methanol, the increase is 20fold at pH 7.4 and 9fold at pH 9.0
Bacillus methanolicus
1.1.1.244
additional information
the enzyme is activated by several Nudix hydrolases, relative activities of Adh enzymes, overview
Bacillus methanolicus
1.1.1.244
NudBC
a Nudix hydrolase from Bacillus coagulans, UniProt ID G2TKZ3
Bacillus methanolicus
1.1.1.244
NudDK
a Nudix hydrolase from Desulfotomaculum kuznetsovii, UniProt ID F6CJI1
Bacillus methanolicus
1.1.1.244
NudLF
a Nudix hydrolase from Lysinibacillus fusiformis, UniProt ID D7WXY3
Bacillus methanolicus
1.1.1.244
NudLS
a Nudix hydrolase from Lysinibacillus sphaericus, UniProt ID B1HRL7
Bacillus methanolicus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.244
gene mdh, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Bacillus methanolicus
1.1.1.244
gene mdh2, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Bacillus methanolicus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.244
NAD+
-
Bacillus methanolicus
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
1.1.1.244
D38G
site-directed mutagenesis, the mutant is active with NADP+ and NAD+ in contrast to the wild-type enzyme
Bacillus methanolicus
1.1.1.244
D41G
site-directed mutagenesis, the mutant is active with NADP+ and NAD+ in contrast to the wild-type enzyme, it shows increased activity compared to the wild-type enzyme
Bacillus methanolicus
1.1.1.244
S101G
site-directed mutagenesis, the mutant shows altered kinetics, reduced activity, and altered pH-dependency compared to the wild-type enzyme, overview
Bacillus methanolicus
1.1.1.244
S98G
site-directed mutagenesis, the mutant shows altered kinetics, reduced activity, and altered pH-dependency compared to the wild-type enzyme, overview
Bacillus methanolicus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.244
additional information
-
additional information
Michaelis-Menten kinetics
Bacillus methanolicus
1.1.1.244
0.009
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.025
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.051
-
ethanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.096
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.137
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in presence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.15
-
methanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.161
-
ethanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.224
-
ethanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.225
-
ethanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.255
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.349
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.379
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in absence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.416
-
methanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.733
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.847
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
1.151
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1.1.1.244
methanol + NAD+
Bacillus methanolicus
-
formaldehyde + NADH + H+
-
-
r
1.1.1.244
methanol + NAD+
Bacillus methanolicus MGA3
-
formaldehyde + NADH + H+
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.244
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Bacillus methanolicus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.1.1.244
ethanol + NAD+
-
742514
Bacillus methanolicus
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.244
ethanol + NAD+
-
742514
Bacillus methanolicus MGA3
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NAD+
-
742514
Bacillus methanolicus
formaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NAD+
-
742514
Bacillus methanolicus MGA3
formaldehyde + NADH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus MGA3
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
methanol + NADP+
Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H)
742514
Bacillus methanolicus MGA3
formaldehyde + NADPH + H+
-
-
-
r
1.1.1.244
additional information
poor activity with 2-propanol and 1,2-popanediol
742514
Bacillus methanolicus
?
-
-
-
-
1.1.1.244
additional information
poor activity with 2-propanol and 1,2-popanediol
742514
Bacillus methanolicus MGA3
?
-
-
-
-
1.1.1.244
n-butanol + NAD+
-
742514
Bacillus methanolicus
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.244
n-butanol + NAD+
-
742514
Bacillus methanolicus MGA3
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.244
n-propanol + NAD+
-
742514
Bacillus methanolicus
propionaldehyde + NADH + H+
-
-
-
r
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.1.244
30
-
assay at
Bacillus methanolicus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.244
9
-
-
Bacillus methanolicus
General Information
EC Number
General Information
Commentary
Organism
1.1.1.244
evolution
the enzyme belongs to the type III alcohol dehydrogenase (Adh) family; the enzyme belongs to the type III alcohol dehydrogenase (Adh) family
Bacillus methanolicus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.244
evolution
the enzyme belongs to the type III alcohol dehydrogenase (Adh) family
Bacillus methanolicus
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.244
0.00004
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0001
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in absence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.0003
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in absence of activator Act; recombinant enzyme, pH 7.4, 30C, in absence of activator Act; recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0006
-
ethanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0006
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in presence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.0007
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in presence of activator Act; recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0008
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0014
-
ethanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0028
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0068
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0074
-
ethanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0143
-
ethanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0353
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
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
1.1.1.244
0.00004
-
methanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0001
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in absence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.0003
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in absence of activator Act; recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0003
-
methanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0006
-
ethanol
recombinant enzyme, pH 7.4, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0006
-
methanol
recombinant enzyme mutant D38G, pH 7.4, 30C, in presence of activator Act, with NADP+
Bacillus methanolicus
1.1.1.244
0.0007
-
methanol
recombinant enzyme mutant S98G, pH 7.4, 30C, in presence of activator Act; recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0008
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0014
-
ethanol
recombinant enzyme, pH 9.0, 30C, in absence of activator Act
Bacillus methanolicus
1.1.1.244
0.0028
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0068
-
methanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0074
-
ethanol
recombinant enzyme, pH 7.4, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0143
-
ethanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus
1.1.1.244
0.0353
-
methanol
recombinant enzyme, pH 9.0, 30C, in presence of activator Act
Bacillus methanolicus