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Literature summary for 1.14.14.1 extracted from

  • Liu, L.; Schmid, R.D.; Urlacher, V.B.
    Cloning, expression, and characterization of a self-sufficient cytochrome P450 monooxygenase from Rhodococcus ruber DSM 44319 (2006), Appl. Microbiol. Biotechnol., 72, 876-882.
    View publication on PubMed
Search for more literature for 1.14.14.1

Application

Application Comment Organism
additional information the Cyt P450 monooxygenases can be utilized in the bioremediation of pollutants, as these enzymes convert chemically inert compounds to more water-soluble, hydroxylated derivatives, which may be suitable substrates for many other enzymes Rhodococcus ruber

Cloned(Commentary)

Cloned (Comment) Organism
DNA sequence and genomic organization determination and analysis, usage of directional genome walking method, overexpression of His6-tagged enzyme in Escherichia coli starin BL21(DE3) Rhodococcus ruber

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0034
-
 NADPH pH 7.4, 25°C Rhodococcus ruber
0.126
-
 NADH pH 7.4, 25°C Rhodococcus ruber

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ heme iron, a cytochrome P450 enzyme Rhodococcus ruber

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
89000
-
 x * 89000, recombinant His6-tagged enzyme, SDS-PAGE Rhodococcus ruber

Organism

Organism UniProt Comment Textmining
Rhodococcus ruber Q52TE7 strain DSM 44319
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by metal affinity chromatography Rhodococcus ruber

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7-ethoxycoumarin + [reduced NADPH-hemoprotein reductase] + O2 O-dealkylation Rhodococcus ruber 7-hydroxycoumarin + [oxidized NADPH-hemoprotein reductase] + H2O + ?
-
 ?
acenaphthene + [reduced NADPH-hemoprotein reductase] + O2
-
 Rhodococcus ruber 1-acenaphthenol + [oxidized NADPH-hemoprotein reductase] + H2O
-
 ?
cytochrome c + NADH + H+ + O2
-
 Rhodococcus ruber ?
-
 r
cytochrome c + O2 + NADH
-
 Rhodococcus ruber ?
-
 r
ethylbenzene + [reduced NADPH-hemoprotein reductase] + O2 recombinant enzyme Rhodococcus ruber 1-phenylethyl alcohol + 2-phenylethyl alcohol + [reduced NADPH-hemoprotein reductase] + O2
-
 ?
fluorene + [reduced NADPH-hemoprotein reductase] + O2
-
 Rhodococcus ruber 9-fluorenol + [oxidized NADPH-hemoprotein reductase] + H2O
-
 ?
indene + [reduced NADPH-hemoprotein reductase] + O2
-
 Rhodococcus ruber indenol + [oxidized NADPH-hemoprotein reductase] + H2O
-
 ?
m-xylene + [reduced NADPH-hemoprotein reductase] + O2 recombinant enzyme Rhodococcus ruber 3-methylbenzyl alcohol + [oxidized NADPH-hemoprotein reductase] + H2O
-
 ?
additional information product identifications, no activity with alpha-pinene, terpeneol, ethoxyresorufin, and cyclohexane, a natural self-sufficient fusion protein consisting of ferredoxin, flavin-containing reductase, and P450 monooxygenase, substrate specificity: in the presence of NADPH, the enzyme shows hydroxylation activity towards polycyclic aromatic hydrocarbons naphthalene, indene, acenaphthene, toluene, fluorene, m-xylene, and ethyl benzene, the conversion of naphthalene, acenaphthene, and fluorene results in respective ring monohydroxylated metabolites, alkyl aromatics like toluene, m-xylene, and ethyl benzene are hydroxylated exclusively at the side chains Rhodococcus ruber ?
-
 ?
naphthalene + [reduced NADPH-hemoprotein reductase] + O2 regioselective oxidation Rhodococcus ruber 1-naphthol + H2O + [oxidized NADPH-hemoprotein reductase] + H2O
-
 ?
toluene + O2 + NAD(P)H + cytochrome c
-
 Rhodococcus ruber benzyl alcohol + H2O + NAD(P)+ + reduced cytochrome c
-
 ?

Subunits

Subunits Comment Organism
? x * 89000, recombinant His6-tagged enzyme, SDS-PAGE Rhodococcus ruber

Synonyms

Synonyms Comment Organism
class IV cytochrome P450 monooxygenase
-
 Rhodococcus ruber

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22 25 assay at Rhodococcus ruber

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6
-
 NADH pH 7.4, 25°C Rhodococcus ruber
12.75
-
 NADPH pH 7.4, 25°C Rhodococcus ruber

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Rhodococcus ruber

Cofactor

Cofactor Comment Organism Structure
cytochrome c
-
 Rhodococcus ruber
Ferredoxin bound Rhodococcus ruber
FMN bound, quantification of the molar ratio of FMN to protein of 1.1:1 Rhodococcus ruber
NAD(P)H NADPH is preferred to NADH giving twofold higher activity Rhodococcus ruber