Activating Compound | Comment | Organism | Structure |
---|---|---|---|
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 | |
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 | |
additional information | the enzyme is activated by several Nudix hydrolases, relative activities of Adh enzymes, overview | Bacillus methanolicus | |
NudBC | a Nudix hydrolase from Bacillus coagulans, UniProt ID G2TKZ3 | Bacillus methanolicus | |
NudDK | a Nudix hydrolase from Desulfotomaculum kuznetsovii, UniProt ID F6CJI1 | Bacillus methanolicus | |
NudLF | a Nudix hydrolase from Lysinibacillus fusiformis, UniProt ID D7WXY3 | Bacillus methanolicus | |
NudLS | a Nudix hydrolase from Lysinibacillus sphaericus, UniProt ID B1HRL7 | Bacillus methanolicus |
Cloned (Comment) | Organism |
---|---|
gene mdh, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus methanolicus |
gene mdh2, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus methanolicus |
Protein Variants | Comment | Organism |
---|---|---|
D38G | site-directed mutagenesis, the mutant is active with NADP+ and NAD+ in contrast to the wild-type enzyme | Bacillus methanolicus |
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 |
S101G | site-directed mutagenesis, the mutant shows altered kinetics, reduced activity, and altered pH-dependency compared to the wild-type enzyme, overview | Bacillus methanolicus |
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Bacillus methanolicus | |
9 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus | |
25 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
51 | - |
ethanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
96 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus | |
137 | - |
methanol | recombinant enzyme mutant D38G, pH 7.4, 30°C, in presence of activator Act, with NADP+ | Bacillus methanolicus | |
150 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
161 | - |
ethanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus | |
224 | - |
ethanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
225 | - |
ethanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
255 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
349 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
379 | - |
methanol | recombinant enzyme mutant D38G, pH 7.4, 30°C, in absence of activator Act, with NADP+ | Bacillus methanolicus | |
416 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
733 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
847 | - |
methanol | recombinant enzyme mutant S98G, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
1151 | - |
methanol | recombinant enzyme mutant S98G, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
methanol + NAD+ | Bacillus methanolicus | - |
formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | Bacillus methanolicus MGA3 | - |
formaldehyde + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus methanolicus | I3DTM5 | - |
- |
Bacillus methanolicus | I3E2P9 | - |
- |
Bacillus methanolicus MGA3 | I3DTM5 | - |
- |
Bacillus methanolicus MGA3 | I3E2P9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Bacillus methanolicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ethanol + NAD+ | - |
Bacillus methanolicus | acetaldehyde + NADH + H+ | - |
r | |
ethanol + NAD+ | - |
Bacillus methanolicus MGA3 | acetaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Bacillus methanolicus | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Bacillus methanolicus MGA3 | formaldehyde + NADH + H+ | - |
r | |
methanol + NADP+ | Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H) | Bacillus methanolicus | formaldehyde + NADPH + H+ | - |
r | |
methanol + NADP+ | Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H) | Bacillus methanolicus | formaldehyde + NADPH + H+ | - |
r | |
methanol + NADP+ | Mdh enzyme mutant D98G, no activity of the wild-type enzyme with NADP(H) | Bacillus methanolicus MGA3 | formaldehyde + NADPH + H+ | - |
r | |
methanol + NADP+ | Mdh enzyme mutant D41G, no activity of the wild-type enzyme with NADP(H) | Bacillus methanolicus MGA3 | formaldehyde + NADPH + H+ | - |
r | |
additional information | poor activity with 2-propanol and 1,2-popanediol | Bacillus methanolicus | ? | - |
? | |
additional information | poor activity with 2-propanol and 1,2-popanediol | Bacillus methanolicus MGA3 | ? | - |
? | |
n-butanol + NAD+ | - |
Bacillus methanolicus | butyraldehyde + NADH + H+ | - |
r | |
n-butanol + NAD+ | - |
Bacillus methanolicus MGA3 | butyraldehyde + NADH + H+ | - |
r | |
n-propanol + NAD+ | - |
Bacillus methanolicus | propionaldehyde + NADH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
MDH | - |
Bacillus methanolicus |
MDH2 | - |
Bacillus methanolicus |
NAD-dependent methanol dehydrogenase | - |
Bacillus methanolicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bacillus methanolicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Bacillus methanolicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Bacillus methanolicus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the type III alcohol dehydrogenase (Adh) family | Bacillus methanolicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00004 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0001 | - |
methanol | recombinant enzyme mutant D38G, pH 7.4, 30°C, in absence of activator Act, with NADP+ | Bacillus methanolicus | |
0.0003 | - |
methanol | recombinant enzyme mutant S98G, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0003 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0003 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0006 | - |
methanol | recombinant enzyme mutant D38G, pH 7.4, 30°C, in presence of activator Act, with NADP+ | Bacillus methanolicus | |
0.0006 | - |
ethanol | recombinant enzyme, pH 7.4, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0007 | - |
methanol | recombinant enzyme mutant S98G, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0007 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0008 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0014 | - |
ethanol | recombinant enzyme, pH 9.0, 30°C, in absence of activator Act | Bacillus methanolicus | |
0.0028 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0068 | - |
methanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0074 | - |
ethanol | recombinant enzyme, pH 7.4, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0143 | - |
ethanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus | |
0.0353 | - |
methanol | recombinant enzyme, pH 9.0, 30°C, in presence of activator Act | Bacillus methanolicus |