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Literature summary extracted from

  • Jia, B.; Park, S.C.; Lee, S.; Pham, B.P.; Yu, R.; Le, T.L.; Han, S.W.; Yang, J.K.; Choi, M.S., Baumeister, W.; Cheong, G.W.
    Hexameric ring structure of a thermophilic archaeon NADH oxidase that produces predominantly H2O (2008), FEBS J., 275, 5355-5366.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.6.3.2 expression in Escherichia coli Thermococcus profundus

Protein Variants

EC Number Protein Variants Comment Organism
1.6.3.2 C122A mutant contains tightly bound FAD. Mutant has similar NADH and NADPH oxidase activity to that of the wild-type protein Thermococcus profundus
1.6.3.2 C45A mutant contains tightly bound FAD. Mutant has less than 10% of the NAD(P)H oxidase activity of the wild-type protein. Mutation alters the reaction to produce H2O2 instead of H2O Thermococcus profundus
1.6.3.2 C45A/C122A mutant contains tightly bound FAD. Mutant has less than 10% of the NAD(P)H oxidase activity of the wild-type protein. Mutation alters the reaction to produce H2O2 instead of H2O Thermococcus profundus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.6.3.2 0.012
-
 NADPH pH 7.2, 75°C Thermococcus profundus
1.6.3.2 0.053
-
 NADH pH 7.2, 75°C Thermococcus profundus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.6.3.2 49000
-
 6 * 49000, ring-shaped hexameric form, SDS-PAGE Thermococcus profundus
1.6.3.2 300000
-
 gel filtration Thermococcus profundus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.6.3.2 2 NAD(P)H + H+ + O2 Thermococcus profundus in anaerobes, flavin-dependent NAD(P)H oxidases play an important role protecting organisms from oxidative stress NAD(P)+ + 2 H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.6.3.2 Thermococcus profundus B2G3S1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.6.3.2 wild-type and mutant enzymes Thermococcus profundus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.3.2 2 NAD(P)H + H+ + O2 in anaerobes, flavin-dependent NAD(P)H oxidases play an important role protecting organisms from oxidative stress Thermococcus profundus NAD(P)+ + 2 H2O
-
 ?
1.6.3.2 2 NADH + H+ + O2 predominantly converts O2 to H2O, but not to H2O2. When NADPH oxidation is performed at 80°C, approximately 2% of the NADPH supplied is used to produce H2O2. Cys45 participates in the direct four-electron reduction of O2 to H2O, and the Cys45 mutation alters the reaction to produce H2O2 instead of H2O. NADPH is more efficient as electron donor compared to NADH Thermococcus profundus NAD+ + 2 H2O
-
 ?
1.6.3.2 2 NADPH + 2 H+ + O2 predominantly converts O2 to H2O. When NADPH oxidation is performed at 80°C, approximately 7% of the NADPH supplied is used to produce H2O2. Cys45 participates in the direct four-electron reduction of O2 to H2O, and the Cys45 mutation alters the reaction to produce H2O2 instead of H2O. NADPH is more efficient as electron donor compared to NADH Thermococcus profundus 2 NADP+ + 2 H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.6.3.2 hexamer 6 * 49000, ring-shaped hexameric form, SDS-PAGE Thermococcus profundus

Synonyms

EC Number Synonyms Comment Organism
1.6.3.2 NOXtp
-
 Thermococcus profundus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.6.3.2 70
-
-
 Thermococcus profundus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.6.3.2 50 70 50°C: about 40% of maximal activity, 70°C: about 70% of maximal activity Thermococcus profundus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.6.3.2 2.5
-
 NADPH pH 7.2, 75°C Thermococcus profundus
1.6.3.2 6.2
-
 NADH pH 7.2, 75°C Thermococcus profundus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.6.3.2 7.5 8
-
 Thermococcus profundus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.6.3.2 6.5 8.5 pH 6.5: about 70% of maximal activity, pH 8.5: about 45% of maximal activity Thermococcus profundus

Cofactor

EC Number Cofactor Comment Organism Structure
1.6.3.2 FAD contains 0.7-0.9 mol FAD per subunit Thermococcus profundus
1.6.3.2 NADH NADPH is more efficient as electron donor compared to NADH Thermococcus profundus
1.6.3.2 NADPH NADPH is more efficient as electron donor compared to NADH Thermococcus profundus

General Information

EC Number General Information Comment Organism
1.6.3.2 physiological function in anaerobes, flavin-dependent NAD(P)H oxidases play an important role protecting organisms from oxidative stress Thermococcus profundus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.6.3.2 116.9
-
 NADH pH 7.2, 75°C Thermococcus profundus
1.6.3.2 206.6
-
 NADPH pH 7.2, 75°C Thermococcus profundus