Cloned (Comment) | Organism |
---|---|
sequence comparisons and phylogenetic analysis, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus tropicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the enzyme-substrate reaction kinetic studies show the allosteric nature of the enzyme and follow pre-steady state using NADH as a co-substrate. Steady-state kinetics | Bacillus tropicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates | Bacillus tropicus | |
Fe2+ | activates by 63.3% | Bacillus tropicus | |
Fe3+ | the catalytic active site of the enzyme has a Fe metal ion-binding centre in contact with residues His124, Glu125, His129 and Glu214. Fe3+ does not activate the enzyme activity | Bacillus tropicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
pentachlorophenol + NADH + H+ + O2 | Bacillus tropicus | - |
2,3,5,6-tetrachloro-1,4-benzoquinone + NADP+ + chloride + H2O | - |
? | |
pentachlorophenol + NADH + H+ + O2 | Bacillus tropicus AOA-CPS1 | - |
2,3,5,6-tetrachloro-1,4-benzoquinone + NADP+ + chloride + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus tropicus | - |
- |
- |
Bacillus tropicus AOA-CPS1 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Bacillus tropicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | enzyme CpsB from Bacillus tropicus reacts with PCP to trap 1,4-tetrachlorobenzoquinone (Tet-CBQ) formed as 2,3,5,6-tetrakis[(2-hydroxyethyl)thio]-1,4-hydroquinone (THTH) | Bacillus tropicus | ? | - |
- |
|
additional information | enzyme CpsB from Bacillus tropicus reacts with PCP to trap 1,4-tetrachlorobenzoquinone (Tet-CBQ) formed as 2,3,5,6-tetrakis[(2-hydroxyethyl)thio]-1,4-hydroquinone (THTH) | Bacillus tropicus AOA-CPS1 | ? | - |
- |
|
pentachlorophenol + NADH + H+ + O2 | - |
Bacillus tropicus | 2,3,5,6-tetrachloro-1,4-benzoquinone + NADP+ + chloride + H2O | - |
? | |
pentachlorophenol + NADH + H+ + O2 | - |
Bacillus tropicus AOA-CPS1 | 2,3,5,6-tetrachloro-1,4-benzoquinone + NADP+ + chloride + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 64000, SDS-PAGE | Bacillus tropicus |
More | the predicted enzyme secondary structure comprises 290 residues of betaalphabeta motifs, 2 beta-sheets, 1 beta-hairpins, 6 strands, 15 alpha-helixes, 24 helix-helix interactions and 25 beta-turns | Bacillus tropicus |
Synonyms | Comment | Organism |
---|---|---|
GM610_18120 | locus name | Bacillus tropicus |
PCP 4-monooxygenase | - |
Bacillus tropicus |
PCP-4-monooxygenase | - |
Bacillus tropicus |
PcpB | - |
Bacillus tropicus |
Phe4MO | - |
Bacillus tropicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Bacillus tropicus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 60 | optimum activity at 30°C, about 60% of maximal activity at 60°C | Bacillus tropicus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 30 | purified enzyme, 180 min, 90% activity remaining | Bacillus tropicus |
40 | - |
purified enzyme, 180 min, 80% activity remaining | Bacillus tropicus |
50 | - |
purified enzyme, 180 min, 65% activity remaining | Bacillus tropicus |
60 | - |
purified enzyme, 180 min, 20% activity remaining | Bacillus tropicus |
70 | - |
purified enzyme, 180 min, no activity remaining | Bacillus tropicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Bacillus tropicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 11.5 | 10% of maximal activity at pH 5.0, 25% of maximal activity at pH 6.0, about 10% of maximal activity at pH 11.5, inactive at at pH 12-13 | Bacillus tropicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Bacillus tropicus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the Bacillus cereus group in the Phe4MO superfamily, sequence comparisons and phylogenetic analysis | Bacillus tropicus |
metabolism | the first step in the PCP degradation pathway is the rate-limiting step involving the hydroxylation of PCP to 1,4-tetrachlorobenzoquinone (Tet-CBQ). The reaction is catalysed by the enzyme PCP 4-monooxygenase (PcpB) in bacteria and cytochrome p450 monooxygenase in fungus. Phe4MO also plays a critical role in xenobiotic compound degradation and lipid metabolism | Bacillus tropicus |
additional information | enzyme homology structure modeling using PDB IDs 4Q3W and 4JPY individually as templates, overview. The catalytic active site of the enzyme has a Fe3+ metal ion-binding centre in contact with residues His124, Glu125, His129 and Glu214 | Bacillus tropicus |
physiological function | efficient degradation of pentachlorophenol (PCP) and involvement of different genes and enzymes in the degradation pathway in Bacillus tropicus strain AOA-CPS1 | Bacillus tropicus |