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Literature summary for 1.6.3.2 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
Search for more literature for 1.6.3.2

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Thermococcus profundus

Protein Variants

Protein Variants Comment Organism
C122A mutant contains tightly bound FAD. Mutant has similar NADH and NADPH oxidase activity to that of the wild-type protein Thermococcus profundus
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
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]

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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

Organism UniProt Comment Textmining
Thermococcus profundus B2G3S1
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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

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

Synonyms

Synonyms Comment Organism
NOXtp
-
 Thermococcus profundus

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 8
-
 Thermococcus profundus

pH Range

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

Cofactor

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

General Information

General Information Comment Organism
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]

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