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Literature summary extracted from

  • Zhang, J.J.; Liu, H.; Xiao, Y.; Zhang, X.E.; Zhou, N.Y.
    Identification and characterization of catabolic para-nitrophenol 4-monooxygenase and para-benzoquinone reductase from Pseudomonas sp. strain WBC-3 (2009), J. Bacteriol., 191, 2703-2710.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.6.5.6 expressed as His-tagged fusion protein in Escherichia coli Pseudomonas sp. WBC-3
1.14.13.166 His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells Pseudomonas sp.
1.14.13.167 His-tagged enzyme is expressed in Escherichia coli BL21(DE3) cells Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.6.5.6 0.0067
-
 p-benzoquinone
-
 Pseudomonas sp. WBC-3
1.6.5.6 0.181
-
 NADPH
-
 Pseudomonas sp. WBC-3
1.14.13.167 0.012
-
 4-nitrophenol His-tagged enzyme, in 20 mM phosphate buffer (pH 7.0), temperature not specified in the publication Pseudomonas sp.
1.14.13.167 0.1374
-
 NADPH His-tagged enzyme, in 20 mM phosphate buffer (pH 7.0), temperature not specified in the publication Pseudomonas sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.13.166 additional information addition of Mg2+ or Mn2+ ions has no effect on the activity Pseudomonas sp.
1.14.13.167 additional information addition of Mg2+ or Mn2+ ions has no effect on the activity Pseudomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.6.5.6 25000
-
 SDS-PAGE Pseudomonas sp. WBC-3
1.14.13.166 47000
-
 1 * 47000, His-tagged enzyme, gel filtration Pseudomonas sp.
1.14.13.167 47000
-
 1 * 47000, His-tagged enzyme, gel filtration Pseudomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
1.6.5.6 Pseudomonas sp. WBC-3 C1I202
-
-
1.14.13.166 Pseudomonas sp. C1I201
-
-
1.14.13.167 Pseudomonas sp. C1I201
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.6.5.6 using Ni2+-NTA affinity chromatography Pseudomonas sp. WBC-3
1.14.13.166 Ni-NTA column chromatography Pseudomonas sp.
1.14.13.167 Ni-NTA column chromatography Pseudomonas sp.

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.14.13.166 culture condition:4-nitrophenol-grown cell
-
 Pseudomonas sp.
-
1.14.13.167 culture condition:4-nitrophenol-grown cell
-
 Pseudomonas sp.
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.166 1.9
-
 at pH 7.0 and 30°C Pseudomonas sp.
1.14.13.167 0.4
-
 crude extract, at pH 7.0 and 30°C Pseudomonas sp.
1.14.13.167 6.8
-
 purified enzyme, at pH 7.0 and 30°C Pseudomonas sp.

Storage Stability

EC Number Storage Stability Organism
1.6.5.6 -70°C, 50% glycerol, 1 month, no loss of activity Pseudomonas sp. WBC-3
1.6.5.6 0°, on ice, 12 h, fairly stable Pseudomonas sp. WBC-3
1.14.13.166 -20°C, without glycerol, rapid inactivation Pseudomonas sp.
1.14.13.166 -70°C, in 50% (v/v) glycerol, 7 days, 70% loss of activity Pseudomonas sp.
1.14.13.166 0°C, storage medium, no loss of activity after several hours, 30% loss of activity after 12 h, 80% loss of activity after 40 h Pseudomonas sp.
1.14.13.167 -20°C, without glycerol, rapid inactivation Pseudomonas sp.
1.14.13.167 -70°C, in 50% (v/v) glycerol, 7 days, 70% loss of activity Pseudomonas sp.
1.14.13.167 0°C, storage medium, no loss of activity after several hours, 30% loss of activity after 12 h, 80% loss of activity after 40 h Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.5.6 NADPH + H+ + p-benzoquinone
-
 Pseudomonas sp. WBC-3 NADP+ + hydroquinone
-
 ?
1.14.13.166 4-nitrocatechol + NADPH + H+ + O2
-
 Pseudomonas sp. ?
-
 ?
1.14.13.167 4-nitrocatechol + NADPH + H+ + O2 lower activity compared to 4-nitrophenol Pseudomonas sp. ?
-
 ?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2 monooxygenation Pseudomonas sp. 4-benzoquinone + nitrite + NADP+ + H2O
-
 ?
1.14.13.167 additional information no activity with 3-nitrophenol, 2-nitrophenol, 4-nitrobenzoate, nitrobenzene, 3-dinitrobenzene, 2,6-dinitrotoluene, 3-hydroxy-4-nitrobenzoate, 2,4,6-trinitrophenol, and 4-chlorophenol Pseudomonas sp. ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.6.5.6 dimer SDS-PAGE, 2 * 25000 Da Pseudomonas sp. WBC-3
1.14.13.166 monomer 1 * 47000, His-tagged enzyme, gel filtration Pseudomonas sp.
1.14.13.167 monomer 1 * 47000, His-tagged enzyme, gel filtration Pseudomonas sp.

Synonyms

EC Number Synonyms Comment Organism
1.6.5.6 p-Benzoquinone reductase
-
 Pseudomonas sp. WBC-3
1.6.5.6 PnpB
-
 Pseudomonas sp. WBC-3
1.14.13.166 para-nitrophenol 4-monooxygenase
-
 Pseudomonas sp.
1.14.13.166 PNP 4-monooxygenase
-
 Pseudomonas sp.
1.14.13.166 pnpA
-
 Pseudomonas sp.
1.14.13.167 para-nitrophenol 4-monooxygenase
-
 Pseudomonas sp.
1.14.13.167 PNP 4-monooxygenase
-
 Pseudomonas sp.
1.14.13.167 pnpA
-
 Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.6.5.6 7.2
-
 assay at Pseudomonas sp. WBC-3

Cofactor

EC Number Cofactor Comment Organism Structure
1.6.5.6 FAD FMN is preferred to FAD Pseudomonas sp. WBC-3
1.6.5.6 FMN enzyme is flavin mononucleotide-dependent, FMN is preferred to FAD Pseudomonas sp. WBC-3
1.6.5.6 NADPH enzyme is NADPH-dependent Pseudomonas sp. WBC-3
1.14.13.166 FAD dependent on, negligible activity is observed when FMN replaces FAD Pseudomonas sp.
1.14.13.166 NADH NADPH can be replaced by NADH with the same efficiency Pseudomonas sp.
1.14.13.166 NADPH
-
 Pseudomonas sp.
1.14.13.167 FAD dependent on, negligible activity is observed when FMN replaces FAD Pseudomonas sp.
1.14.13.167 NADH NADPH can be replaced by NADH with the same efficiency Pseudomonas sp.
1.14.13.167 NADPH
-
 Pseudomonas sp.

Expression

EC Number Organism Comment Expression
1.14.13.167 Pseudomonas sp. 0.0018 mg/ml of para-benzoquinone reductase PnpB enhances the 4-nitrophenol degradation activity by para-nitrophenol 4-monooxygenase by about 2fold up

General Information

EC Number General Information Comment Organism
1.6.5.6 physiological function PnpB enhances the p-nitrophenol degradation by p-nitrophenol 4-monooxygenase Pseudomonas sp. WBC-3
1.14.13.167 physiological function para-nitrophenol 4-monooxygenase gene pnpA plays an essential role in 4-nitrophenol mineralization in strain Pseudomonas putida strain WBC-3 Pseudomonas sp.