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Literature summary for 1.14.13.167 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)

Cloned (Comment) Organism
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.
gene pnpA1, recombinant expression of N-terminally His6-tagged enzyme PnpA in Escherichia coli strain Rosetta(DE3)pLysS Rhodococcus opacus

Protein Variants

Protein Variants Comment Organism
additional information construction of pK18mobsacB-pnpA1 for gene knockout of pnpA1 in strain RKJ300 Rhodococcus opacus
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Burkholderia sp.
-
-
-
Rhodococcus opacus
-
-
-
Rhodococcus opacus RKJ300
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged enzyme PnpA from Escherichia coli strain Rosetta(DE3)pLysS by nickel affinity chromatography Rhodococcus opacus
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]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Burkholderia sp. 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
2-chloro-4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus RKJ300 2-chloro-1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
4-nitrophenol + NADPH + H+ + O2
-
Burkholderia sp. 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
4-nitrophenol + NADPH + H+ + O2
-
Rhodococcus opacus RKJ300 1,4-benzoquinone + nitrite + NADP+ + H2O
-
?
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 ?
-
?
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

Synonyms Comment Organism
2C4NP 4-monooxygenase
-
Rhodococcus opacus
2C4NP 4-monooxygenase
-
Burkholderia sp.
PNP 4-monooxygenase
-
Rhodococcus opacus
PNP 4-monooxygenase
-
Burkholderia sp.
pnpA
-
Burkholderia sp.
PnpA1
-
Rhodococcus opacus

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
FAD
-
Rhodococcus opacus
FAD
-
Burkholderia sp.
NADPH
-
Rhodococcus opacus
NADPH
-
Burkholderia sp.

Expression

Organism Comment Expression
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

General Information Comment Organism
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.
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.
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