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Literature summary for 1.14.15.1 extracted from

  • Hoffmann, G.; Boensch, K.; Greiner-Stoeffele, T.; Ballschmiter, M.
    Changing the substrate specificity of P450cam towards diphenylmethane by semi-rational enzyme engineering (2011), Protein Eng. Des. Sel., 24, 439-446.
    View publication on PubMed
Search for more literature for 1.14.15.1

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type enzyme, encoded by gene camC, with gene camB, encoding the putidaredoxin, in Escherichia coli strains BL21(DE3), expression of mutant enzymes in Escherichia coli strain DH10B Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
I396A site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
I396G site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
I396V site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
M395I site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
M96Y site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
additional information establishment of an in vitro screening system for P450cam selecting variants by the activity on NADH Pseudomonas putida
N244L site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
T297D site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida
V87F site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(1R)-camphor + putidaredoxin + O2 Pseudomonas putida putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
(1R)-camphor + putidaredoxin + O2 Pseudomonas putida PpG1 putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
 gene camC
-
Pseudomonas putida PpG1
-
 gene camC
-

Source Tissue

Source Tissue Comment Organism Textmining
additional information cells grown in D-camphor minimal medium Pseudomonas putida
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method Pseudomonas putida 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
(1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam Pseudomonas putida 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
(1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method Pseudomonas putida PpG1 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
(1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam Pseudomonas putida PpG1 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
additional information substrate specificities of wild-type and mutant enzymes, overview Pseudomonas putida ?
-
 ?
additional information substrate specificities of wild-type and mutant enzymes, overview Pseudomonas putida PpG1 ?
-
 ?

Synonyms

Synonyms Comment Organism
CYP101
-
 Pseudomonas putida
P450cam
-
 Pseudomonas putida
P450cam monooxygenase
-
 Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
putidaredoxin transferring electrons from NADH to P450cam in a coupled assay method Pseudomonas putida