EC Number | Application | Comment | Organism |
---|---|---|---|
1.14.15.1 | additional information | P450cam is not an antifungal target, but is an excellent model in which to study the phenomenon of drug resistance | Pseudomonas putida |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.15.1 | expression of wild-type and mutant enzymes in Escherichia coli | Pseudomonas putida |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.14.15.1 | recombinant wild-type enzyme and mutant L244A/C334A in complex with imidazole or 1-methylimidazole, hanging drop vapour diffusion method, purified recombinant protein in 50 mM potassium phosphate, 250 mM KCl, 50 mM DTT, and 36-52% ammonium sulfate, mixing with an equal volume of 0.001 ml of mother liquor containing 25 mM imidazole or 1-methylimidazole, room temperature, 2 days, cryoprotectant is 50 mM KCl, 25% ammonium sulfate and 30% glycerol, X-ray diffraction structure determination and analysis at 1.5-2.15 A resolution | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.14.15.1 | L244A/C334A | site-directed mutagenesis, mutation C334A prevents adventitious dimerization to facilitate crystallization but has no further effect on structure or activity of the enzyme, the L244A mutation leads to a highly increased Km and reduced activity for imidazole, but not for for 1-methylimidazole, and altered binding of imidazole to the active site and the active site heme involving residue Val247, overview | Pseudomonas putida |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.14.15.1 | 0.0013 | - |
(+)-Camphor | recombinant wild-type enzyme | Pseudomonas putida | |
1.14.15.1 | 0.0016 | - |
(+)-Camphor | recombinant mutant L244A/C334A | Pseudomonas putida | |
1.14.15.1 | 0.0024 | - |
1-Methylimidazole | recombinant wild-type enzyme | Pseudomonas putida | |
1.14.15.1 | 0.0075 | - |
imidazole | recombinant wild-type enzyme | Pseudomonas putida | |
1.14.15.1 | 0.0559 | - |
1-Methylimidazole | recombinant mutant L244A/C334A | Pseudomonas putida | |
1.14.15.1 | 0.3 | - |
imidazole | recombinant mutant L244A/C334A | Pseudomonas putida |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.14.15.1 | Iron | heme iron | Pseudomonas putida |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.15.1 | (+)-camphor + O2 + reduced putidaredoxin | Pseudomonas putida | - |
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.15.1 | Pseudomonas putida | P00183 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.14.15.1 | recombinant wild-type and mutant enzymes from Escherichia coli by anion exchange chromatography and gel filtration to homogeneity | Pseudomonas putida |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.15.1 | (+)-camphor + O2 + reduced putidaredoxin | - |
Pseudomonas putida | (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? | |
1.14.15.1 | 1-methylimidazole + O2 + reduced putidaredoxin | - |
Pseudomonas putida | ? | - |
? | |
1.14.15.1 | imidazole + O2 + reduced putidaredoxin | - |
Pseudomonas putida | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.15.1 | cytochrome P450(cam) | - |
Pseudomonas putida |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.15.1 | putidaredoxin | - |
Pseudomonas putida |