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Literature summary for 1.14.99.15 extracted from

  • Bernhardt, F.H.; Pachowsky, H.; Staudinger, H.
    A 4-methoxybenzoate O-demethylase from Pseudomonas putida. A new type of monooxygenase system (1975), Eur. J. Biochem., 57, 241-256.
    View publication on PubMed
Search for more literature for 1.14.99.15

Activating Compound

Activating Compound Comment Organism Structure
Putidamonooxin
-
 Pseudomonas putida

General Stability

General Stability Organism
ethanol, 5-15% v/v, stabilizes activity in buffer and crude extract Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline
-
 Pseudomonas putida
2,2'-dipyridyl
-
 Pseudomonas putida
5,5'-dithiobis(2-nitrobenzoate)
-
 Pseudomonas putida
8-hydroxyquinoline
-
 Pseudomonas putida
Amytal
-
 Pseudomonas putida
Atebrin
-
 Pseudomonas putida
Bathocuproinedisulfonate
-
 Pseudomonas putida
bathophenanthrolinedisulfate
-
 Pseudomonas putida
diethyldithiocarbamate
-
 Pseudomonas putida
EDTA
-
 Pseudomonas putida
KCN
-
 Pseudomonas putida
p-chloromercuribenzoate reversible by GSH Pseudomonas putida
rotenone
-
 Pseudomonas putida
Thenoyl trifluoroacetone
-
 Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0007
-
 3-Nitro-4-methoxybenzoate
-
 Pseudomonas putida
0.0014
-
 4-Methoxybenzoate
-
 Pseudomonas putida
0.008
-
 NADH reductase Pseudomonas putida
0.077
-
 N-methyl-4-aminobenzoate
-
 Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ both enzyme components are iron-sulfur proteins Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-methoxybenzoate + NADH + O2 Pseudomonas putida
-
 4-hydroxybenzoate + NAD+ + H2O + formaldehyde
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-

Oxidation Stability

Oxidation Stability Organism
putidamonooxin and NADH-reductase, extremely O2-sensitive, purification and storage of the reductase under anaerobic conditions Pseudomonas putida

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein 30% carbohydrate in each of the 50000 Da subunits Pseudomonas putida

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
10.97
-
-
 Pseudomonas putida
21.96
-
 4-methoxybenzoate Pseudomonas putida

Storage Stability

Storage Stability Organism
-20°C, freeze-dried partially purified reductase stable for months without loss of activity, purified reductase is 3-4 weeks stable after addition of NADH in N2-atmosphere Pseudomonas putida
4°C, concentrated putidamonooxin stable for several months in N2-atmosphere with DTT and dithionite Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-nitro-4-methoxybenzoate + NADH + O2
-
 Pseudomonas putida 3-nitro-4-hydroxybenzoate + NAD+ + H2O + formaldehyde
-
 ?
4-methoxybenzoate + NADH + O2
-
 Pseudomonas putida 4-hydroxybenzoate + NAD+ + H2O + formaldehyde
-
 ?
N-methyl-4-aminobenzoate + NADH + O2
-
 Pseudomonas putida 4-aminobenzoate + NAD+ + H2O + formaldehyde
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Pseudomonas putida

pH Range

pH Minimum pH Maximum Comment Organism
7.3 9 about half-maximal activity at pH 7.3 and 9.0, reductase Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
FMN
-
 Pseudomonas putida
NADH
-
 Pseudomonas putida