Literature summary extracted from

Bell, S.G.; Xu, F.; Forward, I.; Bartlam, M.; Rao, Z.; Wong, L.L.
Crystal structure of CYP199A2, a para-substituted benzoic acid oxidizing cytochrome P450 from Rhodopseudomonas palustris (2008), J. Mol. Biol., 383, 561-574.

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
1.14.99.15	overexpression of the His-tagged enzyme in Escherichia coli BL21(DE3)	Rhodopseudomonas palustris

Crystallization (Commentary)

EC Number	Crystallization (Comment)	Organism
1.14.99.15	purified recombinant CYP199A2, 16 ºC, the hanging drop vapor diffusion method under aerobic conditions, 0.0015 ml of protein solution is mixed with 0.0015 ml of reservoir solution, addition of 200 ml reservoir solution, containing 15% PEG 4000, 100 mM sodium citrate pH 5.6, 20% isopropanol with 4% v/v t-butanol, 1 week, X-ray diffraction structure deternnation and analysis at 2.0 A resolution	Rhodopseudomonas palustris

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.14.99.15	additional information	-	additional information	steady-state kinetics	Rhodopseudomonas palustris

Metals/Ions

EC Number	Metals/Ions	Comment	Organism	Structure
1.14.99.15	Fe2+	heme iron	Rhodopseudomonas palustris

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
1.14.99.15	4-methoxybenzoate + reduced ferredoxin + O2	Rhodopseudomonas palustris	-	4-hydroxybenzoate + formaldehyde + ferredoxin + H2O	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
1.14.99.15	Rhodopseudomonas palustris	-	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
1.14.99.15	recombinant His-tagged enzyme from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration	Rhodopseudomonas palustris

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
1.14.99.15	4-ethylbenzoate + AH2 + O2	a C-C bond dehydrogenation of an unbranched alkyl group, computational docking of 4-ethylbenzoate into the active site suggests that the substrate carboxylate oxygens interact with Ser97 and Ser247, and the beta-methyl group is located over the heme iron by Phe185, this binding orientation is consistent with the observed product profile of exclusive attack at the para substituent, overview	Rhodopseudomonas palustris	4-(1-hydroxyethyl)-benzoate + 4-vinylbenzoate + A + H2O	-	?
1.14.99.15	4-methoxybenzoate + reduced ferredoxin + O2	-	Rhodopseudomonas palustris	4-hydroxybenzoate + formaldehyde + ferredoxin + H2O	-	?
1.14.99.15	4-methoxybenzoate + reduced putidaredoxin + O2	very low activity with putidaredoxin	Rhodopseudomonas palustris	4-hydroxybenzoate + formaldehyde + oxidized putidaredoxin + H2O	-	?
1.14.99.15	additional information	CYP199A2 shows a strong preference for para-substituted benzoate over identically substituted ortho- and meta- benzoates, and para-substituted benzenes, benzyl alcohols and benzaldehydes, a cytochrome P450 enzyme, the substrate binding pocket is hydrophobic, with Ser97 and Ser247 being the only polar residues, substrate binding and substrate channeling mechanism and structure, overview	Rhodopseudomonas palustris	?	-	?

Synonyms

EC Number	Synonyms	Comment	Organism
1.14.99.15	CYP199A2	-	Rhodopseudomonas palustris

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
1.14.99.15	30	-	assay at	Rhodopseudomonas palustris

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
1.14.99.15	7.4	-	assay at	Rhodopseudomonas palustris

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
1.14.99.15	reduced ferredoxin	-	Rhodopseudomonas palustris

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Release 2023.1 (January 2023)