Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Putidamonooxin | essential part of the enzyme system, O2-activating 2Fe-2S-protein, identified by EPR and Mössbauer spectroscopy | Pseudomonas putida |
General Stability | Organism |
---|---|
4-methoxybenzoate, stabilizes | Pseudomonas putida |
ethanol, 5-15% v/v, stabilizes activity in buffer and crude extract | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00063 | - |
NADH | reconstituted enzyme | Pseudomonas putida | |
0.03 | - |
Putidamonooxin | - |
Pseudomonas putida | |
0.14 | - |
NADPH | reconstituted enzyme | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | mononuclear non-heme iron protein, 2Fe-2S cluster | Pseudomonas putida | |
Fe2+ | both enzyme components are iron-sulfur proteins | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
41500 | - |
3-4 * 41500, putidamonooxin, SDS-PAGE | Pseudomonas putida |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
- |
- |
Oxidation Stability | Organism |
---|---|
aerobic conditions, 0-4°C, 50% activity lost within 24 h | Pseudomonas putida |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas putida |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-methoxybenzoate + reduced acceptor + O2 = 4-hydroxybenzoate + formaldehyde + acceptor + H2O | the bacterial enzyme is a two-component enzyme, consisting of an iron-sulfur flavoprotein (FMN), NADH-putidamonooxin-reductase and a ferredoxin-type, oxygen-activating protein, putidamonooxin | Pseudomonas putida |
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 |
0-4°C, more than 24 days stable in crude extract under anaerobic conditions | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3,4-dimethoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 4-hydroxy-3-methoxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
3,4-methylenedioxybenzoate + NADH + O2 | piperonylate | Pseudomonas putida | 3,4-dihydroxybenzoate + NAD+ + H2O + formaldehyde | protocatechuate | ? | |
3-hydroxybenzoate + NADH + O2 | partial uncoupler | Pseudomonas putida | 3,4-dihydroxybenzoate + NAD+ + H2O | in the uncoupled part of the reaction, 3-hydroxybenzoate is not hydroxylated and H2O2 is a product, too | ? | |
3-methoxybenzoate + NADH + O2 | partial uncoupler | Pseudomonas putida | 3-hydroxybenzoate + NAD+ + H2O + formaldehyde | in the uncoupled part of the reaction 3-methoxybenzoate is not hydroxylated, and H2O2 is a product, too | ? | |
3-nitro-4-methoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 3-nitro-4-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
3-phenyl-4-methoxybenzoate + NADH + O2 | partial uncoupler | Pseudomonas putida | 3-phenyl-4-hydroxybenzoate + NAD+ + H2O + formaldehyde | in the uncoupled part of the reaction 3-phenyl-4-methoxybenzoate is not hydroxylated and H2O2 is also a product of the reaction | ? | |
4-aminobenzoate + NADH + O2 | - |
Pseudomonas putida | 4-amino-3-hydroxybenzoate + NAD+ + H2O | - |
? | |
4-ethoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 4-hydroxybenzoate + acetaldehyde + NAD+ + H2O | - |
? | |
4-Hydroxy-3-methoxybenzoate + NADH + O2 | vanillate, partial uncoupler | Pseudomonas putida | ? | - |
? | |
4-hydroxybenzoate + NADH + O2 | - |
Pseudomonas putida | 3,4-dihydroxybenzoate + NAD+ + H2O | - |
? | |
4-methoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 4-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
4-methoxybenzoate + NADPH + O2 | - |
Pseudomonas putida | 4-hydroxybenzoate + NADP+ + H2O + formaldehyde | - |
r | |
4-methylbenzoate + NADH + O2 | p-toluate, partial uncoupler | Pseudomonas putida | 4-carboxybenzylalcohol + NAD+ + H2O | in the uncoupled part of the reaction, p-toluate is not hydroxylated and H2O2 is a product, too | ? | |
4-methylmercaptobenzoate + NADH + O2 | - |
Pseudomonas putida | ? | - |
? | |
4-vinylbenzoate + NADH + O2 | external dioxygenase reaction by substrate induced modulation | Pseudomonas putida | 4-glycylbenzoate + NAD+ + H2O | - |
? | |
additional information | overview: substrates that are not oxygenized while NADH-oxidation and O2-consumption are catalyzed, such as: benzoate, 3-chlorobenzoate, 4-chlorobenzoate, 2-hydroxybenzoate, 4-bromobenzoate, 2-aminobenzoate, 3-aminobenzoate, 4-trifluoromethylbenzoate, 4-tert-butylbenzoate | Pseudomonas putida | additional information | uncoupling substrates are not oxygenized, NAD+ and H2O2 being the only products of the reaction | ? | |
additional information | overview: substrates being absolutely planar aromatic rings with a directly bound dissociable carboxy group are oxygenated under stoichiometric consumption of O2 and NADH | Pseudomonas putida | additional information | uncoupling substrates are not oxygenized, NAD+ and H2O2 being the only products of the reaction | ? | |
N,N'-dimethyl-4-aminobenzoate + NADH + O2 | - |
Pseudomonas putida | 4-aminobenzoate + NAD+ + H2O + formaldehyde | - |
? | |
N-methyl-4-aminobenzoate + NADH + O2 | - |
Pseudomonas putida | 4-aminobenzoate + NAD+ + H2O + formaldehyde | - |
? |
Subunits | Comment | Organism |
---|---|---|
oligomer | 3-4 * 41500, putidamonooxin, SDS-PAGE | Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | - |
Pseudomonas putida | |
NADH | requirement, can be replaced by NADPH with 40% efficiency | Pseudomonas putida | |
NADPH | can replace NADH with 40% efficiency | Pseudomonas putida |