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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.1 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

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.1 (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
-
?
1.14.15.1 (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

EC Number Organism UniProt Comment Textmining
1.14.15.1 Pseudomonas putida
-
gene camC
-
1.14.15.1 Pseudomonas putida PpG1
-
gene camC
-

Source Tissue

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.1 (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
-
?
1.14.15.1 (1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam Pseudomonas putida 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
?
1.14.15.1 (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
-
?
1.14.15.1 (1R)-camphor + putidaredoxin + O2 putidaredoxin transfers electrons from NADH to P450cam Pseudomonas putida PpG1 5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
?
1.14.15.1 additional information substrate specificities of wild-type and mutant enzymes, overview Pseudomonas putida ?
-
?
1.14.15.1 additional information substrate specificities of wild-type and mutant enzymes, overview Pseudomonas putida PpG1 ?
-
?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.1 CYP101
-
Pseudomonas putida
1.14.15.1 P450cam
-
Pseudomonas putida
1.14.15.1 P450cam monooxygenase
-
Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.15.1 30
-
assay at Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.15.1 7.4
-
assay at Pseudomonas putida

Cofactor

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