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Literature summary for 1.6.5.5 extracted from

  • Reis, R.A.G.; Salvi, F.; Williams, I.; Gadda, G.
    Kinetic investigation of a presumed nitronate monooxygenase from Pseudomonas aeruginosa PAO1 establishes a new class of NAD(P)H quinone reductases (2019), Biochemistry, 58, 2594-2607 .
    View publication on PubMed
Search for more literature for 1.6.5.5

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta(DE3)pLysS cells Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
additional information when NADH is used instead of NADPH, substrate inhibition is not observed Pseudomonas aeruginosa
NADPH substrate inhibition is observed at NADPH concentrations of more than 0.05 mM Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.012
-
 NADPH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.016
-
 NADPH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.074
-
 5-hydroxy-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.1
-
 1,4-Naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.23
-
 2-methyl-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.23
-
 NADH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.24
-
 NADH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.37
-
 1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.5
-
 2,3-dimethoxy-5-methyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.5
-
 2,3,4,6-tetramethyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
2.5
-
 2,6-dimethoxy-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9I5R1
-
-

Purification (Commentary)

Purification (Comment) Organism
HiTrap column chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,4-benzoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 1,4-benzoquinol + NADP+
-
 ?
1,4-naphthoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 1,4-naphthoquinol + NADP+
-
 ?
2,3,4,6-tetramethyl-1,4-benzoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 2,3,4,6-tetramethyl-1,4-benzoquinol + NADP+
-
 ?
2,3-dimethoxy-5-methyl-1,4-benzoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 2,3-dimethoxy-5-methyl-1,4-benzoquinol + NADP+
-
 ?
2,6-dichlorophenolindophenol + NADH + H+ the enzyme is 25times more specific for NADPH as a substrate than for NADH Pseudomonas aeruginosa reduced 2,6-dichlorophenolindophenol + NAD+
-
 ?
2,6-dichlorophenolindophenol + NADPH + H+ the enzyme is 25times more specific for NADPH as a substrate than for NADH Pseudomonas aeruginosa reduced 2,6-dichlorophenolindophenol + NADP+
-
 ?
2,6-dimethoxy-1,4-benzoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 2,6-dimethoxy-1,4-benzoquinol + NADP+
-
 ?
2-methyl-1,4-naphthoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 2-methyl-1,4-naphthoquinol + NADP+
-
 ?
5-hydroxy-1,4-naphthoquinone + NADPH + H+
-
 Pseudomonas aeruginosa 5-hydroxy-1,4-naphthoquinol + NADP+
-
 ?
additional information the enzyme does not exhibit nitronate monooxygenase activity (no activity with propionate 3-nitronate, 3-nitropropionate, nitroethane, 2-nitropropane, ethylnitronate, or propyl-2-nitronate) and does not turn over with methyl red, consistent with a lack of azoreductase activity Pseudomonas aeruginosa ?
-
-
O2 + NADH + H+ the enzyme displays negligible NAD(P)H:oxidase activity Pseudomonas aeruginosa ?
-
 ?
O2 + NADPH + H+ the enzyme displays negligible NAD(P)H:oxidase activity Pseudomonas aeruginosa ?
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 72000, analytical ultracentrifugation Pseudomonas aeruginosa
homodimer 2 * 72200, calculated from amino acid sequence Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
NAD(P)H:quinone reductase
-
 Pseudomonas aeruginosa
PA0660
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 NADH with O2 cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.12
-
 NADPH with O2 cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
12
-
 NADH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
13
-
 NADH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
15
-
 2,3,4,6-tetramethyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
15
-
 NADPH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
16
-
 NADPH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
41
-
 2,6-dimethoxy-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
63
-
 2-methyl-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
66
-
 2,3-dimethoxy-5-methyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
80
-
 1,4-Naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
82
-
 5-hydroxy-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
87
-
 1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
90
-
 1,4-benzoquinone at pH 7.5 and 25°C Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
FMN
-
 Pseudomonas aeruginosa
NADH the enzyme can be reduced with NADPH or NADH with a marked preference for NADPH Pseudomonas aeruginosa
NADPH the enzyme can be reduced with NADPH or NADH with a marked preference for NADPH Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.17
-
 NADPH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
0.2
-
 NADPH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
16
-
 2,6-dimethoxy-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
30
-
 2,3,4,6-tetramethyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
50
-
 NADH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
50
-
 NADH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
130
-
 2,3-dimethoxy-5-methyl-1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
250
-
 1,4-benzoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
300
-
 2-methyl-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
1000
-
 NADPH with 0.08 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
1100
-
 5-hydroxy-1,4-naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
1250
-
 1,4-Naphthoquinone with NADPH as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa
1300
-
 NADPH with 0.06 mM 2,6-dichlorophenolindophenol as cosubstrate, at pH 7.5 and 25°C Pseudomonas aeruginosa