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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Burkholderia sp. | - |
- |
- |
Rhodococcus opacus | - |
- |
- |
Rhodococcus opacus RKJ300 | - |
- |
- |
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 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 | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Rhodococcus opacus |
30 | - |
assay at | Burkholderia sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Rhodococcus opacus |
7.4 | - |
assay at | Burkholderia sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Rhodococcus opacus | |
FAD | - |
Burkholderia sp. | |
NADPH | - |
Rhodococcus opacus | |
NADPH | - |
Burkholderia sp. |
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 | 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 |