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Literature summary for 1.1.2.8 extracted from

  • Good, N.M.; Vu, H.N.; Suriano, C.J.; Subuyuj, G.A.; Skovran, E.; Martinez-Gomez, N.C.
    Pyrroloquinoline quinone ethanol dehydrogenase in Methylobacterium extorquens AM1 extends lanthanide-dependent metabolism to multicarbon substrates (2016), J. Bacteriol., 198, 3109-3118 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.2.8

Activating Compound

Activating Compound Comment Organism Structure
ammonium
-
 Methylorubrum extorquens
methylamine methylamine is a 5fold better activator for enzyme ExaF in vitro than ammonium Methylorubrum extorquens

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
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

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ ativates, active site-bound Methylorubrum extorquens
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
additional information the enzyme from strain AM1 utilizes La3+ rather than Ca2+ as a cofactor Methylorubrum extorquens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
126400
-
 recombinant His6-tagged enzyme, gel filtration Methylorubrum extorquens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ethanol + 2 cytochrome c Methylorubrum extorquens
-
 acetaldehyde + 2 reduced cytochrome c
-
 r
ethanol + 2 cytochrome c Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1
-
 acetaldehyde + 2 reduced cytochrome c
-
 r
methanol + 2 cytochrome c Methylorubrum extorquens
-
 formaldehyde + 2 reduced cytochrome c
-
 r
methanol + 2 cytochrome c Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1
-
 formaldehyde + 2 reduced cytochrome c
-
 r
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 ?
-
 ?
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 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Methylorubrum extorquens
-
-
-
Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain TOP10 by ultracentrifugation, nickel affinity chromatography, ultrafiltration, and desalting gel filtration Methylorubrum extorquens

Source Tissue

Source Tissue Comment Organism Textmining
additional information growth rate constants of Methylobacterium extorquens wild-type and mutant strains grown with ethanol as the carbon source Methylorubrum extorquens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ethanol + 2 cytochrome c
-
 Methylorubrum extorquens acetaldehyde + 2 reduced cytochrome c
-
 r
ethanol + 2 cytochrome c
-
 Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 acetaldehyde + 2 reduced cytochrome c
-
 r
methanol + 2 cytochrome c
-
 Methylorubrum extorquens formaldehyde + 2 reduced cytochrome c
-
 r
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
methanol + 2 cytochrome c
-
 Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1 formaldehyde + 2 reduced cytochrome c
-
 r
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
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 ?
-
 ?
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 ?
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 61000, recombinant His6-tagged enzyme, SDS-PAGE Methylorubrum extorquens

Synonyms

Synonyms Comment Organism
exaF
-
 Methylorubrum extorquens
pyrroloquinoline quinone ethanol dehydrogenase
-
 Methylorubrum extorquens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Methylorubrum extorquens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
 assay at, alcohol oxidation Methylorubrum extorquens

Cofactor

Cofactor Comment Organism Structure
cytochrome c
-
 Methylorubrum extorquens
additional information enzyme assays are performed with the artificial electron acceptors phenazine methosulfate and 2,6-dichlorophenolindophenol Methylorubrum extorquens
pyrroloquinoline quinone prosthetic group Methylorubrum extorquens

General Information

General Information Comment Organism
malfunction an exaF mutant is not affected for growth with ethanol Methylorubrum extorquens
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
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