Application | Comment | Organism |
---|---|---|
additional information | P450cam is not an antifungal target, but is an excellent model in which to study the phenomenon of drug resistance | Pseudomonas putida |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
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 |
Protein Variants | Comment | Organism |
---|---|---|
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 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0013 | - |
(+)-Camphor | recombinant wild-type enzyme | Pseudomonas putida | |
0.0016 | - |
(+)-Camphor | recombinant mutant L244A/C334A | Pseudomonas putida | |
0.0024 | - |
1-Methylimidazole | recombinant wild-type enzyme | Pseudomonas putida | |
0.0075 | - |
imidazole | recombinant wild-type enzyme | Pseudomonas putida | |
0.0559 | - |
1-Methylimidazole | recombinant mutant L244A/C334A | Pseudomonas putida | |
0.3 | - |
imidazole | recombinant mutant L244A/C334A | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | heme iron | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-camphor + O2 + reduced putidaredoxin | Pseudomonas putida | - |
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P00183 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli by anion exchange chromatography and gel filtration to homogeneity | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-camphor + O2 + reduced putidaredoxin | - |
Pseudomonas putida | (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? | |
1-methylimidazole + O2 + reduced putidaredoxin | - |
Pseudomonas putida | ? | - |
? | |
imidazole + O2 + reduced putidaredoxin | - |
Pseudomonas putida | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cytochrome P450(cam) | - |
Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
putidaredoxin | - |
Pseudomonas putida |