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Literature summary for 1.14.15.1 extracted from
- The role of putidaredoxin and P450 cam in methylene hydroxylation (1972), J. Biol. Chem., 247, 5777-5784.
General Stability
| General Stability | Organism |
|---|---|
| freeze-thawing, less than 5% loss of activity of cytochrome P450cam in the presence of camphor | Pseudomonas putida |
| putidaredoxin suffers degradation by repeated cycles of freezing and thawing | Pseudomonas putida |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytoplasm | - |
Pseudomonas putida | 5737 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | requirement, enzyme complex with two FeS-protein components | Pseudomonas putida |  |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| additional information | - |
putidaredoxin reductase: MW 43000 Da, putidaredoxin: MW 12500 Da, cytochrome P450cam: MW 45000 Da, Pseudomonas putida PpG786, ultracentrifugal, diffusion and amino acid analysis | Pseudomonas putida |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (+)-camphor + putidaredoxin + O2 | Pseudomonas putida | - |
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | i.e. (+)-exo-5-hydroxycamphor | ? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas putida | - |
PpG786, mutant derived from PpG1 | - |
| Pseudomonas putida | - |
Accession Number PC00183 | - |
Oxidation Stability
| Oxidation Stability | Organism |
|---|---|
| under aerobic conditions, cytochrome P450cam decays at 25°C with t1/2 of 180 min to cytochrome P420, not rapidly in the presence of camphor | Pseudomonas putida |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| PpG786 | Pseudomonas putida |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| (+)-camphor + reduced putidaredoxin + O2 = (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | multi-component mixed function oxidase, consisting of putidaredoxin reductase, putidaredoxin and cytochrome P450cam, heme-thiolate protein | Pseudomonas putida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (+)-camphor + putidaredoxin + O2 | - |
Pseudomonas putida | (R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | i.e. (+)-exo-5-hydroxycamphor | ? | |
| (+)-camphor + putidaredoxin + O2 | the (+)- and (-)-enantiomers serve as substrates | Pseudomonas putida | (R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | i.e. (+)-exo-5-hydroxycamphor | ? | |
| 1,2-campholide + putidaredoxin + O2 | - |
Pseudomonas putida | 5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O | - |
? | |
| additional information | - |
Pseudomonas putida | ? | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Pseudomonas putida |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
assay at | Pseudomonas putida |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| cytochrome m | - |
Pseudomonas putida | |
| FMN | - |
Pseudomonas putida | |
| NADH | requirement, reduces reductase flavoprotein and putidaredoxin, but not P450cam, in the absence of camphor | Pseudomonas putida | |
| putidaredoxin | cannot be replaced by other FeS-proteins or the phospholipid of the hepatic microsomal P450 system | Pseudomonas putida |
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