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Literature summary for 1.2.5.3 extracted from

  • Kaufmann, P.; Duffus, B.R.; Teutloff, C.; Leimkuehler, S.
    Functional Studies on Oligotropha carboxidovorans molybdenum-copper CO dehydrogenase produced in Escherichia coli (2018), Biochemistry, 57, 2889-2901 .
    View publication on PubMed

Application

Application Comment Organism
synthesis establishment of a functional heterologous expression system of Moco-free apo-CODH in Escherichia coli. The expression of the CoxMSL genes alone results in a colorless and inactive protein that lacks the cofactors. Expression of an active protein requires coexpression of CoxI Afipia carboxidovorans

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Afipia carboxidovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00052
-
CO pH 7.2, 30°C Afipia carboxidovorans

Metals/Ions

Metals/Ions Comment Organism Structure
copper
-
Afipia carboxidovorans
Molybdenum
-
Afipia carboxidovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
285000
-
gel filtration Afipia carboxidovorans

Organism

Organism UniProt Comment Textmining
Afipia carboxidovorans P19921 and P19920 and P19919 P19921 i.e. small subunit CoxS, P19920 i.e. medium subunit CoxM, P19919 i.e. large subunit CoxL
-
Afipia carboxidovorans DSM 1227 P19921 and P19920 and P19919 P19921 i.e. small subunit CoxS, P19920 i.e. medium subunit CoxM, P19919 i.e. large subunit CoxL
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein, purification by Ni-NTA and size exclusion chromatography Afipia carboxidovorans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.3
-
recombinant purified enzyme, pH 7.2, 30°C Afipia carboxidovorans
2.6
-
recombinant purified enzyme, reconstitution of the active site with sulfur and copper, pH 7.2, 30°C Afipia carboxidovorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CO + 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride + H2O
-
Afipia carboxidovorans CO2 + reduced 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride
-
?
CO + 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride + H2O
-
Afipia carboxidovorans DSM 1227 CO2 + reduced 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride
-
?
additional information is able to catalyze both the oxidation of CO to CO2 and the oxidation of H2 to protons and electrons Afipia carboxidovorans ?
-
-
additional information is able to catalyze both the oxidation of CO to CO2 and the oxidation of H2 to protons and electrons Afipia carboxidovorans DSM 1227 ?
-
-

Synonyms

Synonyms Comment Organism
CoxS
-
Afipia carboxidovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.6
-
CO pH 7.2, 30°C Afipia carboxidovorans

Cofactor

Cofactor Comment Organism Structure
[2Fe-2S]-center presence of 2 [Fe2S2] clusters, UV-vis spectrum shows a shoulder at 550 nm Afipia carboxidovorans