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Literature summary for 1.14.13.50 extracted from

  • Aregbesola, O.A.; Kumar, A.; Mokoena, M.P.; Olaniran, A.O.
    Classic pentachlorophenol hydroxylating phenylalanine 4-monooxygenase from indigenous Bacillus tropicus strain AOA-CPS1 cloning, overexpression, purification, characterization and structural homology modelling (2022), Appl. Biochem. Biotechnol., 194, 635-658 .
    View publication on PubMed

Cloned(Commentary)

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 [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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Bacillus tropicus
-
-
-
Bacillus tropicus AOA-CPS1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Bacillus tropicus

Substrates and Products (Substrate)

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

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Bacillus tropicus

Temperature Range [°C]

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 [°C]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Bacillus tropicus

pH Range

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

Cofactor Comment Organism Structure
NADH
-
Bacillus tropicus

General Information

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