EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
1.1.2.8 | ammonium | - |
Methylorubrum extorquens | |
1.1.2.8 | methylamine | methylamine is a 5fold better activator for enzyme ExaF in vitro than ammonium | Methylorubrum extorquens |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.2.8 | gene exaF, or exaA, DNA and amino acid sequence determination and analysis, functional recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain TOP10 | Methylorubrum extorquens |
1.1.2.10 | expressed in Escherichia coli Top10 cells | Methylorubrum extorquens |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.2.8 | additional information | a triple mutant strain (mxaF xoxF1 xoxF2, named MDH-3), deficient in the three methanol dehydrogenases of the model methylotroph Methylobacterium extorquens AM1, is able to grow poorly with methanol if exogenous lanthanides are added to the growth medium. When the gene encoding a putative quinoprotein ethanol dehydrogenase, exaF, is mutated in the MDH-3 background, the quadruple mutant strain can no longer grow on methanol in minimal medium with added lanthanum (La3+) | Methylorubrum extorquens |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.2.10 | 0.0009 | - |
ethanol | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 0.066 | - |
formaldehyde | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 0.285 | - |
acetaldehyde | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 5.98 | - |
methanol | at pH 9.0 and 30°C | Methylorubrum extorquens |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.1.2.8 | Ca2+ | ativates, active site-bound | Methylorubrum extorquens | |
1.1.2.8 | La3+ | required, active site-bound, 1.3 mol of La3+ per mol of ExaF protomer, indicating a 1:1 ratio of L3+ to protomer of enzyme | Methylorubrum extorquens | |
1.1.2.8 | additional information | the enzyme from strain AM1 utilizes La3+ rather than Ca2+ as a cofactor | Methylorubrum extorquens | |
1.1.2.10 | La3+ | the enzyme preferentially binds La3+ over Ca2+ in the active site. 0.1 mM used in assay conditions. The enzyme contains 1.3 mol of La3+ per mol of protomer | Methylorubrum extorquens |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.1.2.8 | 126400 | - |
recombinant His6-tagged enzyme, gel filtration | Methylorubrum extorquens |
1.1.2.10 | 126400 | - |
SEC-MALS | Methylorubrum extorquens |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.2.8 | ethanol + 2 cytochrome c | Methylorubrum extorquens | - |
acetaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | ethanol + 2 cytochrome c | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | - |
acetaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | Methylorubrum extorquens | - |
formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | - |
formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | additional information | Methylorubrum extorquens | ExaF is functionally an ethanol dehydrogenase, with secondary activities with formaldehyde, methanol, and acetaldehyde. Enzyme assays are performed with with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol. Methanol-independent reduction of the latter is not observed with purified enzyme | ? | - |
? | |
1.1.2.8 | additional information | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | ExaF is functionally an ethanol dehydrogenase, with secondary activities with formaldehyde, methanol, and acetaldehyde. Enzyme assays are performed with with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol. Methanol-independent reduction of the latter is not observed with purified enzyme | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.2.8 | Methylorubrum extorquens | - |
- |
- |
1.1.2.8 | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | - |
- |
- |
1.1.2.10 | Methylorubrum extorquens | C5AXV8 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.2.8 | recombinant His6-tagged enzyme from Escherichia coli strain TOP10 by ultracentrifugation, nickel affinity chromatography, ultrafiltration, and desalting gel filtration | Methylorubrum extorquens |
1.1.2.10 | Ni-NTA column chromatography | Methylorubrum extorquens |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|---|---|---|---|
1.1.2.8 | additional information | growth rate constants of Methylobacterium extorquens wild-type and mutant strains grown with ethanol as the carbon source | Methylorubrum extorquens | - |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.2.8 | ethanol + 2 cytochrome c | - |
Methylorubrum extorquens | acetaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | ethanol + 2 cytochrome c | - |
Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | acetaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | - |
Methylorubrum extorquens | formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | the maximal reaction velocity of ExaF-La for formaldehyde is 23% greater than for methanol as the substrate | Methylorubrum extorquens | formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | - |
Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | methanol + 2 cytochrome c | the maximal reaction velocity of ExaF-La for formaldehyde is 23% greater than for methanol as the substrate | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | formaldehyde + 2 reduced cytochrome c | - |
r | |
1.1.2.8 | additional information | ExaF is functionally an ethanol dehydrogenase, with secondary activities with formaldehyde, methanol, and acetaldehyde. Enzyme assays are performed with with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol. Methanol-independent reduction of the latter is not observed with purified enzyme | Methylorubrum extorquens | ? | - |
? | |
1.1.2.8 | additional information | ExaF is functionally an ethanol dehydrogenase, with secondary activities with formaldehyde, methanol, and acetaldehyde. Enzyme assays are performed with with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol. Methanol-independent reduction of the latter is not observed with purified enzyme | Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 | ? | - |
? | |
1.1.2.10 | ethanol + phenazine methosulfate | highest activity | Methylorubrum extorquens | acetaldehyde + reduced phenazine methosulfate | - |
r | |
1.1.2.10 | formaldehyde + reduced phenazine methosulfate | the enzyme is 100times more efficient at oxidizing formaldehyde than methanol | Methylorubrum extorquens | methanol + phenazine methosulfate | - |
r | |
1.1.2.10 | methanol + phenazine methosulfate | - |
Methylorubrum extorquens | formaldehyde + reduced phenazine methosulfate | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.2.8 | homodimer | 2 * 61000, recombinant His6-tagged enzyme, SDS-PAGE | Methylorubrum extorquens |
1.1.2.10 | homodimer | x * 61000, SDS-PAGE | Methylorubrum extorquens |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.2.8 | exaF | - |
Methylorubrum extorquens |
1.1.2.8 | pyrroloquinoline quinone ethanol dehydrogenase | - |
Methylorubrum extorquens |
1.1.2.10 | exaF | - |
Methylorubrum extorquens |
1.1.2.10 | pyrroloquinoline quinone ethanol dehydrogenase | - |
Methylorubrum extorquens |
1.1.2.10 | quinoprotein ethanol dehydrogenase | - |
Methylorubrum extorquens |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.2.8 | 30 | - |
assay at | Methylorubrum extorquens |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.2.8 | 9 | - |
assay at, alcohol oxidation | Methylorubrum extorquens |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.2.8 | cytochrome c | - |
Methylorubrum extorquens | |
1.1.2.8 | additional information | enzyme assays are performed with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol | Methylorubrum extorquens | |
1.1.2.8 | pyrroloquinoline quinone | prosthetic group | Methylorubrum extorquens | |
1.1.2.10 | pyrroloquinoline quinone | - |
Methylorubrum extorquens |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.2.8 | malfunction | an exaF mutant is not affected for growth with ethanol | Methylorubrum extorquens |
1.1.2.8 | additional information | ExaF homology modeling using the crystal structure of the quinoprotein ethanol dehydrogenase QEDH from Pseudomonas aeruginosa, PDB 1FLG, overview. Residues E198, D317, D319, and N275 form the active site. Residue D319 might be necessary for lanthanide coordination next to catalytic aspartate D317 | Methylorubrum extorquens |
1.1.2.8 | physiological function | ExaF contributes to ethanol metabolism when La3 is present, expanding the role of lanthanides to multicarbon metabolism. ExaA quinoprotein ethanol dehydrogenase, and not the type I ADH,EC 1.1.2.4, is responsible for methanol oxidation in the MDH-3 mutant strain | Methylorubrum extorquens |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.2.10 | 2 | - |
methanol | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 20 | - |
acetaldehyde | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 259 | - |
formaldehyde | at pH 9.0 and 30°C | Methylorubrum extorquens | |
1.1.2.10 | 14500 | - |
ethanol | at pH 9.0 and 30°C | Methylorubrum extorquens |