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Literature summary extracted from

  • Michizoe, J.; Ichinose, H.; Kamiya, N.; Maruyama, T.; Goto, M.
    Functionalization of the cytochrome P450cam monooxygenase system in the cell-like aqueous compartments of water-in-oil emulsions (2005), J. Biosci. Bioeng., 99, 12-17.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.99.22 expression in Escherichia coli BL21(DE3)pLysS as His-tag fusion protein Escherichia coli
1.14.15.1 individual expression of enzyme and cofactor putidaredoxin in Escherichia coli Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
1.14.15.1 additional information establishment of a functional CYP101-system in a cell-like aqueous compartment build by a stable water-oil emulsion with the non-ioninc surfactant tetraethylene glycol dodecyl ether in micro-scale, the enzyme is not active in an organic-aqueous biphasic system, e.g. with etahnol or glycol, incorporating an NADH-regeneration system using recombinant His-tagged bacterial glycerol dehydrogenase, method optimization, overview 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 (+)-camphor + O2 + reduced putidaredoxin Pseudomonas putida
-
 (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.99.22 Escherichia coli
-
-
-
1.1.99.22 Escherichia coli JM109
-
-
-
1.14.15.1 Pseudomonas putida
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.99.22 to homogeneity by anion exchange chromatography and Ni-affinity chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.99.22 glycerol + NAD+
-
 Escherichia coli dihydroxyacetone + NADH + H+
-
 ?
1.1.99.22 glycerol + NAD+
-
 Escherichia coli JM109 dihydroxyacetone + NADH + H+
-
 ?
1.14.15.1 (+)-camphor + O2 + reduced putidaredoxin
-
 Pseudomonas putida (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?
1.14.15.1 (+)-camphor + O2 + reduced putidaredoxin mechanism of camphor hydroxylation incorporating an NADH-regeneration system, overview Pseudomonas putida (+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.99.22 GLD
-
 Escherichia coli
1.1.99.22 glycerol dehydrogenase
-
 Escherichia coli
1.14.15.1 CYP101
-
 Pseudomonas putida
1.14.15.1 cytochrome P450cam monooxygenase
-
 Pseudomonas putida
1.14.15.1 P450cam
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.15.1 37
-
 assay at Pseudomonas putida

pH Optimum

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

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.1 putidaredoxin
-
 Pseudomonas putida