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Literature summary for 1.14.12.13 extracted from

  • Fetzner, S.; Muller, R.; Lingens, F.
    Purification and some properties of 2-halobenzoate 1,2-dioxygenase, a two-component enzyme system from Pseudomonas cepacia 2CBS (1992), J. Bacteriol., 174, 279-290.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.12.13

Inhibitors

Inhibitors Comment Organism Structure
2,2'-dipyridyl 100% inhibition at 1 mM Burkholderia cepacia
Cu2+ 100% inhibition compared to the activity without any metal Burkholderia cepacia
EDTA 100% inhibition at 1 mM Burkholderia cepacia
iodoacetate 55% inhibition at 2 mM Burkholderia cepacia
KCN 40% inhibition at 2 mM Burkholderia cepacia
additional information no inhibition with sodium azide Burkholderia cepacia
N-ethylmaleimide 100% inhibition at 2 mM Burkholderia cepacia
Ni2+ 25% inhibition compared to the activity without any metal Burkholderia cepacia
o-phenanthroline 100% inhibition at 0.5 mM Burkholderia cepacia
p-chloromercuribenzoate 17% inhibition at 0.005 mM Burkholderia cepacia
Zn2+ 40% inhibition compared to the activity without any metal Burkholderia cepacia

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.023
-
 2-Chlorobenzoate component A Burkholderia cepacia
0.079
-
 NADH component B Burkholderia cepacia

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ exogenous, required for activity Burkholderia cepacia
Fe2+ component B contains 1.7 mol and A 5.8 mol of iron per mol of enzyme Burkholderia cepacia
Sulfide component B contains 1.7 mol and A 6.0 mol of acid-labile sulfide per mol of enzyme Burkholderia cepacia

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
20000
-
 3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE Burkholderia cepacia
37500
-
 1 * 37500, component B, SDS-PAGE Burkholderia cepacia
37500 38000 component B, gel filtration Burkholderia cepacia
52000
-
 3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE Burkholderia cepacia
200000 220000 component A, gel filtration Burkholderia cepacia

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-chlorobenzoate + NADH + H+ + O2 Burkholderia cepacia first step of 2-chlorobenzoate degradation via meta-cleavage catechol + chloride + NAD+ + CO2
-
 ?

Organism

Organism UniProt Comment Textmining
Burkholderia cepacia
-
 2CBS, isolated from water
-

Purification (Commentary)

Purification (Comment) Organism
component A: anion-exchange, gel filtration, component B: anion-exchange, affinity and adsorption chromatography Burkholderia cepacia

Reaction

Reaction Comment Organism Reaction ID
2-chlorobenzoate + NADH + H+ + O2 = catechol + chloride + NAD+ + CO2 inducible two-component enzyme system: component B is an iron-sulfur flavoprotein (reductase) and red-brown component A (oxygenase) is an iron-sulfur protein Burkholderia cepacia
a
2-chlorobenzoate + NADH + H+ + O2 = catechol + chloride + NAD+ + CO2 multi-component dioxygenase system proposed: 1,2-hydroxylating, dehalogenating, decarboxylating Burkholderia cepacia
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0085
-
 crude extract, 3.4 mg protein per ml in the assay Burkholderia cepacia
0.53
-
 activity of component A after gel filtration in presence of component B Burkholderia cepacia
4
-
 activity of component B in presence of component A Burkholderia cepacia

Storage Stability

Storage Stability Organism
-20°C component A, 80-85% loss of activity Burkholderia cepacia
-20°C, component A, 20% glycerol, 50% loss of activity Burkholderia cepacia
-20°C, component B, 20% sucrose, 2 weeks, 25% loss of activity Burkholderia cepacia
-80°C or -20°C, component B, 2 weeks, 70% loss of activity Burkholderia cepacia
-80°C or -20°C, component B, 20% glycerol, 2 weeks, 15% loss of activity Burkholderia cepacia

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-aminobenzoate + NADH + O2
-
 Burkholderia cepacia ? + NAD+ + CO2
-
 ?
2-bromobenzoate + NADH + O2
-
 Burkholderia cepacia catechol + bromide + NAD+ + CO2
-
 ?
2-chlorobenzoate + NADH + H+ + O2
-
 Burkholderia cepacia catechol + chloride + NAD+ + CO2
-
 ?
2-chlorobenzoate + NADH + H+ + O2 first step of 2-chlorobenzoate degradation via meta-cleavage Burkholderia cepacia catechol + chloride + NAD+ + CO2
-
 ?
2-fluorobenzoate + NADH + O2
-
 Burkholderia cepacia catechol + fluoride + NAD+ + CO2
-
 ?
2-hydroxybenzoate + NADH + O2
-
 Burkholderia cepacia catechol + ? + NAD+ + CO2
-
 ?
2-iodobenzoate + NADH + O2
-
 Burkholderia cepacia catechol + iodide + NAD+ + CO2
-
 ?
2-methoxybenzoate + NADH + O2
-
 Burkholderia cepacia catechol + ? + NAD+ + CO2
-
 ?
2-methylbenzoate + NADH + O2
-
 Burkholderia cepacia o-cresol + ? + NAD+ + CO2
-
 ?
anthranilate + NADH + O2
-
 Burkholderia cepacia catechol + ? + NAD+ + CO2
-
 ?
benzoate + NADH + H+ + O2
-
 Burkholderia cepacia catechol + NAD+ + CO2
-
 ?
additional information very broad substrate specificity is listed, benzoate analogs with substituents in ortho position are preferred. Trichloroethylene, dihydroxybenzoates, benzene chlorobenzene, toluene, phenol and chlorophenylacetate isomers do not serve as substrates Burkholderia cepacia ?
-
 ?

Subunits

Subunits Comment Organism
hexamer 3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE Burkholderia cepacia
monomer 1 * 37500, component B, SDS-PAGE Burkholderia cepacia

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Burkholderia cepacia

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Burkholderia cepacia

Cofactor

Cofactor Comment Organism Structure
FAD component B contains 0.8 mol per mol of enzyme, 0.002 mM increases activity Burkholderia cepacia
FMN little increase of activity by addition of 0.002 mM Burkholderia cepacia
NADH can not be replaced by NADPH Burkholderia cepacia