Cloned (Comment) | Organism |
---|---|
pCHC1 plasmid, encoding the wild type cytochrome P450cam (C334A mutant of the native enzyme). PCR products transformed into Escherichia coli XL1-blue super-competent cells. Wild-type and mutants expressed in Escherichia coli BL21 (DE3) cells | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
S190D | does not show any significant change in the rate constants of the substrate association, has almost no effect on the activation energy of substrate binding to the enzyme | Pseudomonas putida |
T192E | rate constants of the substrate association is much lower compared to the wild-type, activation energy for the substrate association is significantly higher in the T192E mutant compared to the S190D mutant or the wild-type enzyme | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P00183 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutants | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-camphor + O2 + reduced putidaredoxin | though the active site of the enzyme resides deep inside the protein matrix, the substrate is recognized at the surface of the enzyme and directed towards the active site through the access channel. The threonine 192 that resides on the F-G loop and directed towards the putative substrate access channel of the enzyme, plays an important role in recognition of the substrate at the surface of the enzyme | Pseudomonas putida | (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cytochrome p450cam | - |
Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
putidaredoxin | - |
Pseudomonas putida |