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Literature summary for 1.14.12.9 extracted from

  • Schweizer, D.; Markus, A.; Seez, M.; Ruf, H.H.; Lingens, F.
    Purification and some properties of component B of the 4-chlorophenylacetate 3,4-dioxygenase from Pseudomonas species strain CBS 3 (1987), J. Biol. Chem., 262, 9340-9346.
    View publication on PubMed
Search for more literature for 1.14.12.9

General Stability

General Stability Organism
75% loss of activity after 16 h dialysis Pseudomonas sp.

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 1 mM, 34% inhibition Pseudomonas sp.
EDTA 1 mM, 32% inhibition Pseudomonas sp.
iodoacetate 0.01 mM, 77% inhibition Pseudomonas sp.
KCN 1 mM, 52% inhibition Pseudomonas sp.
NEM 1 mM, 38% inhibition Pseudomonas sp.
NH4+ inhibition of activity of reductase component Pseudomonas sp.
PCMB 0.005 mM, complete inhibition Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0023
-
 ferrocytochrome c
-
 Pseudomonas sp.
0.0063
-
 reduced 2,6-dichlorophenolindophenol
-
 Pseudomonas sp.
0.032
-
 NADH
-
 Pseudomonas sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Iron reductase component contains an iron-sulfur cluster of the [2Fe-2S]-type, contains 2.1 mol of iron and 1.7 mol of acid-labile sulfide per mol of reductase Pseudomonas sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
 reductase component of 4-chlorophenylacetate 3,4-dioxygenase, gel filtration Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-
Pseudomonas sp. CBS 3
-
-
-

Oxidation Stability

Oxidation Stability Organism
reductase component loses activity in presence of molecular oxygen Pseudomonas sp.

Purification (Commentary)

Purification (Comment) Organism
reductase component Pseudomonas sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
30
-
 reductase component of of 4-chlorophenylacetate 3,4-dioxygenase Pseudomonas sp.

Storage Stability

Storage Stability Organism
4°C, 20 mM potassium phosphate buffer, pH 6.0-7.5, 10 days, 50% loss of activity Pseudomonas sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-chlorophenylacetate + ferrocytochrome c + H+ + O2
-
 Pseudomonas sp. 3,4-dihydroxyphenylacetate + chloride + ferricytochrome c
-
 r
4-chlorophenylacetate + ferrocytochrome c + H+ + O2
-
 Pseudomonas sp. CBS 3 3,4-dihydroxyphenylacetate + chloride + ferricytochrome c
-
 r
4-chlorophenylacetate + NADH + O2 + H2O
-
 Pseudomonas sp. 3,4-dihydroxyphenylacetate + Cl- + NAD(P)+
-
 ?
4-chlorophenylacetate + NADH + O2 + H2O
-
 Pseudomonas sp. CBS 3 3,4-dihydroxyphenylacetate + Cl- + NAD(P)+
-
 ?
4-chlorophenylacetate + reduced 2,6-dichlorophenolindophenol + O2
-
 Pseudomonas sp. 3,4-dihydroxyphenylacetate + chloride + oxidized 2,6-dichlorophenolindophenol
-
 r
4-chlorophenylacetate + reduced 2,6-dichlorophenolindophenol + O2
-
 Pseudomonas sp. CBS 3 3,4-dihydroxyphenylacetate + chloride + oxidized 2,6-dichlorophenolindophenol
-
 r
additional information the reductase component reduces several redox compounds: 2,6-dichlorophenylindophenol, potassium hexacyanoferrate III, cytochrome c, methylene blue and nitro blue tetrazolium Pseudomonas sp. ?
-
 ?
additional information the reductase component reduces several redox compounds: 2,6-dichlorophenylindophenol, potassium hexacyanoferrate III, cytochrome c, methylene blue and nitro blue tetrazolium Pseudomonas sp. CBS 3 ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 35000, reductase component of of 4-chlorophenylacetate 3,4-dioxygenase, SDS-PAGE Pseudomonas sp.
More the enzyme consists of two components, a monomeric reductase and a trimeric dioxygenase Pseudomonas sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 3 min, 50% loss of activity Pseudomonas sp.
55
-
 90-100% loss of activity Pseudomonas sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 7.5
-
 Pseudomonas sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 7.5 4°C, 10 days, 50% loss of activity Pseudomonas sp.

Cofactor

Cofactor Comment Organism Structure
FMN flavoprotein, contains 1 mol of FMN per mol of reductase, no increase of activity by addition of exogenous FMN Pseudomonas sp.
NADH
-
 Pseudomonas sp.
NADPH 30% as effective as NADH Pseudomonas sp.