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Literature summary extracted from

  • Min, J.; Zhang, J.J.; Zhou, N.Y.
    The gene cluster for para-nitrophenol catabolism is responsible for 2-chloro-4-nitrophenol degradation in Burkholderia sp. strain SJ98 (2014), Appl. Environ. Microbiol., 80, 6212-6222 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.167 gene pnpA, expression of the pnp cluster is detected only under inducing conditions, and pnpABA1CDEFG is a single transcriptional operon, genetic structure, RT-PCR and real-time PCR enzyme expression analysis, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS Burkholderia sp.
1.14.13.167 gene pnpA1, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS Rhodococcus opacus

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.167 additional information construction of pK18mobsacB-pnpA1 for gene knockout of pnpA1 in strain RKJ300 Rhodococcus opacus
1.14.13.167 additional information enzyme deletion strain SJ98DELTApnpA1 exhibits specific activities for 2-chloro-4-nitrophenol approximately 20% lower than those of wild-type SJ98 Burkholderia sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.167 additional information
-
additional information Michaelis-Menten kinetics Burkholderia sp.
1.14.13.167 0.0062
-
2-chloro-4-nitrophenol pH 7.4, 30°C, recombinant enzyme Burkholderia sp.
1.14.13.167 0.0254
-
4-nitrophenol pH 7.4, 30°C, recombinant enzyme Burkholderia sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.29 4-nitrophenol + NAD(P)H + H+ + O2 Rhodococcus opacus
-
4-nitrocatechol + NAD(P)+ + H2O
-
?
1.14.13.29 4-nitrophenol + NAD(P)H + H+ + O2 Rhodococcus opacus RKJ300
-
4-nitrocatechol + NAD(P)+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2 Rhodococcus opacus
-
2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2 Burkholderia sp.
-
2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2 Rhodococcus opacus RKJ300
-
2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2 Rhodococcus opacus
-
1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2 Burkholderia sp.
-
1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2 Rhodococcus opacus RKJ300
-
1,4-benzoquinone + nitrite + NADP+ + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.6.5.6 Burkholderia sp. SJ98
-
-
-
1.14.13.29 Rhodococcus opacus
-
-
-
1.14.13.29 Rhodococcus opacus RKJ300
-
-
-
1.14.13.167 Burkholderia sp.
-
-
-
1.14.13.167 Rhodococcus opacus
-
-
-
1.14.13.167 Rhodococcus opacus RKJ300
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.167 recombinant N-terminally His6-tagged enzyme PnpA from Escherichia coli strain Rosetta(DE3)pLysS by nickel affinity chromatography Rhodococcus opacus
1.14.13.167 recombinant N-terminally His6-tagged enzyme PnpA from Escherichia coli strain Rosetta(DE3)pLysS by nickel affinity chromatography Burkholderia sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.167 1.73
-
mutant strain SJ98DELTApnpA1 cell extract, pH 7.5, 30°C, substrate 2-chloro-4-nitrophenol Burkholderia sp.
1.14.13.167 2.1
-
cell extract, pH 7.4, 30°C, substrate 2-chloro-4-nitrophenol Burkholderia sp.
1.14.13.167 3.74
-
mutant strain SJ98DELTApnpA1 cell extract, pH 7.5, 30°C, substrate 4-nitrophenol Burkholderia sp.
1.14.13.167 4.8
-
cell extract, pH 7.4, 30°C, substrate 4-nitrophenol Burkholderia sp.
1.14.13.167 5.6
-
pH 7.4, 30°C, substrate 4-nitrophenol Burkholderia sp.
1.14.13.167 7.2
-
pH 7.4, 30°C, substrate 2-chloro-4-nitrophenol Burkholderia sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.5.6 NADPH + H+ + chloro-1,4-benzoquinone
-
Burkholderia sp. SJ98 NADP+ + chlorohydroquinone
-
?
1.6.5.6 NADPH + H+ + p-benzoquinone
-
Burkholderia sp. SJ98 NADP+ + hydroquinone
-
?
1.14.13.29 4-nitrophenol + NAD(P)H + H+ + O2
-
Rhodococcus opacus 4-nitrocatechol + NAD(P)+ + H2O
-
?
1.14.13.29 4-nitrophenol + NAD(P)H + H+ + O2
-
Rhodococcus opacus RKJ300 4-nitrocatechol + NAD(P)+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Burkholderia sp. 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus RKJ300 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2
-
Burkholderia sp. 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus RKJ300 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
1.14.13.167 additional information PnpA1A2 also convert 2-chloro-1,4-benzoquinone to 2-hydroxy-4-nitrophenol, the combined isozymes also show low activity with chlorohydroquinone forming 2-hydroxy-1,4-hydroquinone, metabolite identification by GC-MS analysis, overview. Isozyme PnpA2 is not active with 2-chloro-4-nitrophenol Rhodococcus opacus ?
-
?
1.14.13.167 additional information PnpA1A2 also convert 2-chloro-1,4-benzoquinone to 2-hydroxy-4-nitrophenol, the combined isozymes also show low activity with chlorohydroquinone forming 2-hydroxy-1,4-hydroquinone, metabolite identification by GC-MS analysis, overview. Isozyme PnpA2 is not active with 2-chloro-4-nitrophenol Rhodococcus opacus RKJ300 ?
-
?

Synonyms

EC Number Synonyms Comment Organism
1.6.5.6 PnpB
-
Burkholderia sp. SJ98
1.14.13.167 2C4NP 4-monooxygenase
-
Rhodococcus opacus
1.14.13.167 2C4NP 4-monooxygenase
-
Burkholderia sp.
1.14.13.167 PNP 4-monooxygenase
-
Rhodococcus opacus
1.14.13.167 PNP 4-monooxygenase
-
Burkholderia sp.
1.14.13.167 pnpA
-
Burkholderia sp.
1.14.13.167 PnpA1
-
Rhodococcus opacus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.167 30
-
assay at Rhodococcus opacus
1.14.13.167 30
-
assay at Burkholderia sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.167 7
-
assay at Rhodococcus opacus
1.14.13.167 7.4
-
assay at Burkholderia sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.29 NAD(P)H
-
Rhodococcus opacus
1.14.13.167 FAD
-
Rhodococcus opacus
1.14.13.167 FAD
-
Burkholderia sp.
1.14.13.167 NADPH
-
Rhodococcus opacus
1.14.13.167 NADPH
-
Burkholderia sp.

Expression

EC Number Organism Comment Expression
1.14.13.167 Burkholderia sp. 2-chloro-4-nitrophenol degradation is induced by either 2-chloro-4-nitrophenol or 4-nitrophenol, negligible activity in uninduced cells up

General Information

EC Number General Information Comment Organism
1.6.5.6 physiological function enzyme is able to enhance 4-nitrophenol 4-monooxygenase PnpA activity in vitro in the conversion of 2-chloro-4-nitrophenol to chloro-1,4-benzoquinone Burkholderia sp. SJ98
1.14.13.167 metabolism the enzymes responsible for 4-nitrophenol catabolism are also likely to be involved in 2-chloro-4-nitrophenol degradation. Catabolism of 2-chloro-4-nitrophenol and of 4-nitrophenolshare the same gene cluster in strain SJ98 Burkholderia sp.
1.14.13.167 physiological function PnpA, a PNP 4-monooxygenase, is able to catalyze the monooxygenation of 2-chloro-4-nitrophenol to 2-chloro-1,4-benzoquinone. PnpB, a 1,4-benzoquinone reductase, has the ability to catalyze the reduction of 2-chloro-1,4-benzoquinone to chlorohydroquinone. Moreover, PnpB is also able to enhance PnpA activity in vitro in the conversion of 2-chloro-4-nitrophenol to 2-chloro-1,4-benzoquinone. Gene pnpA plays an essential role in the degradation of both 2-chloro-4-nitrophenol and 4-nitrophenol. Catabolism of 2-chloro-4-nitrophenol and of 4-nitrophenolshare the same gene cluster in strain SJ98 Burkholderia sp.
1.14.13.167 physiological function PnpA1, a PNP 4-monooxygenase, is able to catalyze the monooxygenation of 2-chloro-4-nitrophenol to 2-chloro-1,4-benzoquinone Rhodococcus opacus