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Literature summary extracted from

  • Wieser, M.; Wagner, B.; Eberspaecher, J.; Lingens, F.
    Purification and characterization of 2,4,6-trichlorophenol-4-monooxygenase, a dehalogenating enzyme from Azotobacter sp. strain GP1 (1997), J. Bacteriol., 179, 202-208 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.14.14.173 additional information ATP (0.01 to 1 mM), ADP (0.01 to 0.1 mM), and coenzyme A (0.5 mM) do not enhance activity of the purified enzyme Cupriavidus sp. GP1

General Stability

EC Number General Stability Organism
1.14.14.173 dithiothreitol (0.5 mM), glycerol (10% (v/v)), sucrose (10% (w/v)), or bovine liver catalase (30 ng/ml), separately or in combinations, slightly enhance enzyme stability. No influence on stability is found for 10% (v/v) ethanol, isopropanol, or H2O2 (0.1 mM). FAD (0.03 mM) leads to a slight destabilization Cupriavidus sp. GP1

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.14.173 2,4,6-Trichlorophenol at 2,4,6-trichlorophenol concentrations higher than 0.1 mM, substrate inhibition which amounts to 43% at 0.2 mM 2,4,6-trichlorophenol and to 72% at 0.33 mM 2,4,6-trichlorophenol is observed Cupriavidus sp. GP1
1.14.14.173 4-hydroxymercuribenzoate 62% inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Acriflavine complete inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Cu2+ complete inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Hg2+ 87% inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 additional information not inhibited by menadione, metapyrone, EDTA, Tiron, 1,10-phenanthroline, KCN and sodium azide (1 mM each) Cupriavidus sp. GP1
1.14.14.173 N-ethylmaleimide 42% inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Quinacrine complete inhibition at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Zn2+ 31% inhibition at 0.1 mM Cupriavidus sp. GP1

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.14.173 Co2+ slight activation at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 Fe2+ slight activation at 0.1 mM. The enzyme contains 0.2 mol of Fe per mol of the homotetrameric protein Cupriavidus sp. GP1
1.14.14.173 Fe3+ slight activation at 0.1 mM Cupriavidus sp. GP1
1.14.14.173 additional information Fe2+ (0.2 mM) and Mg2+ (0.8 mM) do not enhance activity of the purified enzyme. No influence on enzyme activity is observed for 1 mM Mg2+, Ca2+, and Co2+ Cupriavidus sp. GP1
1.14.14.173 Ni2+ slight activation at 0.1 mM Cupriavidus sp. GP1

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.14.173 240000
-
 gel filtration Cupriavidus sp. GP1

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 Cupriavidus sp. GP1
-
 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 Cupriavidus sp. GP1 DSM 6428
-
 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 Cupriavidus sp. GP1 ATCC 49806
-
 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2 Cupriavidus sp. GP1
-
 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2 Cupriavidus sp. GP1 DSM 6428
-
 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2 Cupriavidus sp. GP1 ATCC 49806
-
 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.14.173 Cupriavidus sp. GP1
-
-
-
1.14.14.173 Cupriavidus sp. GP1 ATCC 49806
-
-
-
1.14.14.173 Cupriavidus sp. GP1 DSM 6428
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.173 DEAE-cellulose column chromatography, DE52 cellulose anion-exchange chromatography, and Resource Q anion -exchange chromatography Cupriavidus sp. GP1

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.14.173 0.18
-
 crude extract, pH and temperature not specified in the publication Cupriavidus sp. GP1
1.14.14.173 6.87
-
 after 38.2fold purification, pH and temperature not specified in the publication Cupriavidus sp. GP1

Storage Stability

EC Number Storage Stability Organism
1.14.14.173 -20 C, the purified enzyme is stable for several months Cupriavidus sp. GP1
1.14.14.173 25 C, purified enzyme, 8-15 h, complete loss of activity Cupriavidus sp. GP1
1.14.14.173 4 C, purified enzyme, 2 to 3 days, complete loss of activity Cupriavidus sp. GP1

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.173 2,3,4,5-tetrachlorophenol + FADH2 + O2 33% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,3,4,5-tetrachlorophenol + FADH2 + O2 33% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 DSM 6428 ?
-
 ?
1.14.14.173 2,3,4,5-tetrachlorophenol + FADH2 + O2 33% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ATCC 49806 ?
-
 ?
1.14.14.173 2,3,4,6-tetrachlorophenol + FADH2 + O2 67% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,4,5-trichlorophenol + FADH2 + O2 33% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,4,6-tribromophenol + FADH2 + O2 57% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2
-
 Cupriavidus sp. GP1 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 highest conversion rate Cupriavidus sp. GP1 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2
-
 Cupriavidus sp. GP1 DSM 6428 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 highest conversion rate Cupriavidus sp. GP1 DSM 6428 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2
-
 Cupriavidus sp. GP1 ATCC 49806 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-trichlorophenol + FADH2 + O2 highest conversion rate Cupriavidus sp. GP1 ATCC 49806 2,6-dichloro-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,4,6-triiodophenol + FADH2 + O2 50% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,4-dichlorophenol + FADH2 + O2 80% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2
-
 Cupriavidus sp. GP1 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2
-
 Cupriavidus sp. GP1 DSM 6428 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorohydroquinone + FADH2 + O2
-
 Cupriavidus sp. GP1 ATCC 49806 2-chloro-6-hydroxy-1,4-benzoquinone + Cl- + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorophenol + FADH2 + O2 9% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 2,6-dichlorohydroquinone + FAD + H2O
-
 ?
1.14.14.173 2,6-dichlorophenol + FADH2 + O2 67% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 2-chlorophenol + FADH2 + O2 25% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 3,4,5-trichlorophenol + FADH2 + O2 25% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 3,4-dichlorophenol + FADH2 + O2 50% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 4-chlorophenol + FADH2 + O2 32% activity compared to 2,4,6-trichlorophenol Cupriavidus sp. GP1 ?
-
 ?
1.14.14.173 additional information no activity with 3-chlorophenol, 2,3-dichlorophenol, 2,5-dichlorophenol, 3,5-dichlorophenol, 2,3,4-trichlorophenol, 2,3,5-trichlorophenol, 2,3,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, pentachlorophenol, 2,4,5-trimethylphenol, and 2,4,6-trinitrophenol Cupriavidus sp. GP1 ?
-
-
1.14.14.173 additional information no activity with 3-chlorophenol, 2,3-dichlorophenol, 2,5-dichlorophenol, 3,5-dichlorophenol, 2,3,4-trichlorophenol, 2,3,5-trichlorophenol, 2,3,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, pentachlorophenol, 2,4,5-trimethylphenol, and 2,4,6-trinitrophenol Cupriavidus sp. GP1 DSM 6428 ?
-
-
1.14.14.173 additional information no activity with 3-chlorophenol, 2,3-dichlorophenol, 2,5-dichlorophenol, 3,5-dichlorophenol, 2,3,4-trichlorophenol, 2,3,5-trichlorophenol, 2,3,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, pentachlorophenol, 2,4,5-trimethylphenol, and 2,4,6-trinitrophenol Cupriavidus sp. GP1 ATCC 49806 ?
-
-

Subunits

EC Number Subunits Comment Organism
1.14.14.173 homotetramer 4 * 60000, SDS-PAGE Cupriavidus sp. GP1

Synonyms

EC Number Synonyms Comment Organism
1.14.14.173 2,4,6-trichlorophenol-4-monooxygenase
-
 Cupriavidus sp. GP1
1.14.14.173 TCP-4-monooxygenase
-
 Cupriavidus sp. GP1

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.173 35 40
-
 Cupriavidus sp. GP1

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.173 7
-
 optimum activity in 25 to 50 mM Tris-HCl buffer, pH 7.0 Cupriavidus sp. GP1

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.173 FAD 0.03 mM FAD leads to a 4-5fold activation of the DEAE-cellulose-purified enzyme. FAD cannot be replaced by flavin mononucleotide or riboflavin Cupriavidus sp. GP1
1.14.14.173 NADH the activity with NADH is about 20fold higher than with NADPH Cupriavidus sp. GP1