EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.15.1 | open reading frames for P450cam mutants including a 6 x His carboxyl-terminal tag cloned into the pCW(Ori+) expression vector using the NdeI and XbaI restriction sites, expressed in Escherichia coli | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.14.15.1 | T252A | non-productive H2O2 generation is dominant, does not oxidize camphor, substrate binding affinity is similar to that of the wild-type enzyme | Pseudomonas putida |
1.14.15.1 | T252N | has comparable turnover number but higher Km value relative to the wild-type enzyme, due to a decrease in the camphor binding affinity, non-productive H2O2 generation is negligible | Pseudomonas putida |
1.14.15.1 | T252N/V253T | has comparable turnover number but higher Km value relative to the wild-type enzyme, due to a decrease in the camphor binding affinity, non-productive H2O2 generation is negligible | Pseudomonas putida |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.14.15.1 | 0.03 | - |
(+)-Camphor | wild-type | Pseudomonas putida | |
1.14.15.1 | 0.096 | - |
(+)-Camphor | mutant T252N | Pseudomonas putida | |
1.14.15.1 | 0.388 | - |
(+)-Camphor | mutant T252N/V253T | Pseudomonas putida |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.14.15.1 | Fe2+ | Asn252 can stabilize the ferric hydroperoxy intermediate, preventing premature release of H2O2 and enabling addition of the second proton to the distal oxygen to generate the catalytic ferryl species | Pseudomonas putida |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.14.15.1 | 47000 | - |
gel filtration | Pseudomonas putida |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.15.1 | Pseudomonas putida | P00183 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.14.15.1 | soluble proteins separated by ultracentrifugation and purified using Ni2+-nitrilotriacetate column and gel filtration | 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 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.15.1 | CYP101 | - |
Pseudomonas putida |
1.14.15.1 | cytochrome p450cam | - |
Pseudomonas putida |
1.14.15.1 | P450cam | - |
Pseudomonas putida |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.14.15.1 | 0.9 | - |
(+)-Camphor | mutant T252N | Pseudomonas putida | |
1.14.15.1 | 2.067 | - |
(+)-Camphor | wild-type | Pseudomonas putida | |
1.14.15.1 | 2.75 | - |
(+)-Camphor | mutant T252N/V253T | Pseudomonas putida |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.14.15.1 | putidaredoxin | - |
Pseudomonas putida |