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Literature summary for 1.14.15.1 extracted from

  • Kim, D.; Heo, Y.S.; Ortiz de Montellano, P.R.
    Efficient catalytic turnover of cytochrome P450(cam) is supported by a T252N mutation (2008), Arch. Biochem. Biophys., 474, 150-156.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.15.1

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
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
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
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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
 (+)-Camphor wild-type Pseudomonas putida
0.096
-
 (+)-Camphor mutant T252N Pseudomonas putida
0.388
-
 (+)-Camphor mutant T252N/V253T Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
47000
-
 gel filtration Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P00183
-
-

Purification (Commentary)

Purification (Comment) Organism
soluble proteins separated by ultracentrifugation and purified using Ni2+-nitrilotriacetate column and gel filtration Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(+)-camphor + O2 + reduced putidaredoxin
-
 Pseudomonas putida (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?

Synonyms

Synonyms Comment Organism
CYP101
-
 Pseudomonas putida
cytochrome p450cam
-
 Pseudomonas putida
P450cam
-
 Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.9
-
 (+)-Camphor mutant T252N Pseudomonas putida
2.067
-
 (+)-Camphor wild-type Pseudomonas putida
2.75
-
 (+)-Camphor mutant T252N/V253T Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
putidaredoxin
-
 Pseudomonas putida