Literature summary for 1.14.15.1 extracted from

Johnson, E.O.; Wong, L.L.
Partial fusion of a cytochrome P450 system by carboxy-terminal attachment of putidaredoxin reductase to P450cam (CYP101A1) (2016), Catal. Sci. Technol., 6, 7549-7560 .

Search for more literature for 1.14.15.1

Application

Application	Comment	Organism
synthesis	fusion of putidaredoxin reductase PdR to the carboxy-terminus of camphor monooxygenase CYP101A1 (P450cam) via a linker peptide and reconstitution of camphor hydroxylase activity with free putidaredoxin enables the production of the almost fully heme-incorporated CYP-FdR fusion that is catalytically active in vivo and in vitro	Pseudomonas putida

Cloned(Commentary)

Cloned (Comment)	Organism
gene camC, recombinant expression of wild-type enzyme and chimeric enzyme fusion protein P450cam-PdR in Escherichia coli strain BL21 (DE3)	Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment)	Organism
fusion of putidaredoxin reductase PdR to the carboxy-terminus of camphor monooxygenase CYP101A1 (P450cam) via a linker peptide and reconstitution of camphor hydroxylase activity with free putidaredoxin gives a functional system with comparable in vivo camphor oxidation activity as the native system. In vitro, the fused systems steady state NADH oxidation rate is 2fold faster than that of the native system. In contrast to the native system, NADH oxidation rates for the fusion enzyme show nonhyperbolic dependence on putidaredoxin concentration	Pseudomonas putida

Crystallization (Comment)

Organism

fusion of putidaredoxin reductase PdR to the carboxy-terminus of camphor monooxygenase CYP101A1 (P450cam) via a linker peptide and reconstitution of camphor hydroxylase activity with free putidaredoxin gives a functional system with comparable in vivo camphor oxidation activity as the native system. In vitro, the fused systems steady state NADH oxidation rate is 2fold faster than that of the native system. In contrast to the native system, NADH oxidation rates for the fusion enzyme show nonhyperbolic dependence on putidaredoxin concentration

Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of a fusion enzyme composed of enzyme P450cam and putidaroxin reductase, i.e. P450cam-PdR, kinetic model based on two-site binding of putidaredoxin by P450cam-PdR and inactive dimer formation of the fusion protein. Fusion co-expression with putidaredoxin results in a functional system with in vivo camphor oxidation activity comparable to the wild-type system , but P450cam-PdR is a class I P450 fusion protein that exhibits significantly more favorable catalytic behavior than that of the wild-type system. Further oxidation of 5-exo-hydroxycamphor to 5-oxo-camphor by the fusion enzyme is 39% lower than for the native system	Pseudomonas putida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	[2Fe-2S] putidaredoxin	Pseudomonas putida

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	P00183	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type enzyme and chimeric enzyme fusion protein P450cam-PdR from Escherichia coli strain BL21 (DE3)	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(+)-camphor + O2 + reduced putidaredoxin	-	Pseudomonas putida	exo-5-hydroxycamphor + oxidized putidaredoxin + H2O	-	?
(+)-camphor + reduced putidaredoxin + O2	-	Pseudomonas putida	(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O	-	?
DL-camphor + reduced putidaredoxin + NADH + H+ + O2	-	Pseudomonas putida	exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O	-	?
additional information	comparison of substrate binding and catalytic ability of wild-type enzyme with putidareductase and putidaredoxin (ternary complex), and recombinant fusion enzyme P450cam-putidareductase with putidaredoxin, overview. The wild-type and mutant systems show comparable activity with camphor. Further oxidation of 5-exo-hydroxycamphor to 5-oxo-camphor by the fusion enzyme is 39% lower than for the native system	Pseudomonas putida	?	-	?

Synonyms

Synonyms	Comment	Organism
CamC	-	Pseudomonas putida
camphor 5-hydroxylase	-	Pseudomonas putida
class I cytochrome P450	-	Pseudomonas putida
CYP101A1	-	Pseudomonas putida
P450cam	-	Pseudomonas putida

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
56.4	-	reduced putidaredoxin	putidaredoxin reductase-camphor monooxygenase fusion enzyme, pH 7.4, 30°C	Pseudomonas putida
64	-	reduced putidaredoxin	wild-type, pH 7.4, 30°C	Pseudomonas putida

pH Optimum

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

Cofactor

Cofactor	Comment	Organism
cytochrome P450	-	Pseudomonas putida
cytochrome P450	the putidaredoxin reductase-camphor monooxygenase fusion enzyme shows the 450 nm Soret band characteristic of the CYP superfamily	Pseudomonas putida
NADH	-	Pseudomonas putida
putidaredoxin	[2Fe-2S] putidaredoxin, Pdx. Camphor-bound ferric P450cam domain forms a complex with Pdx, modeling	Pseudomonas putida