Literature summary for 1.11.2.4 extracted from

Matsunaga, I.; Ueda, A.; Sumimoto, T.; Ichihara, K.; Ayata, M.; Ogura, H.
Site-directed mutagenesis of the putative distal helix of peroxygenase cytochrome P450 (2001), Arch. Biochem. Biophys., 394, 45-53.

Search for more literature for 1.11.2.4

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli M15 (pREP4) cells	Bacillus subtilis

Protein Variants

Protein Variants	Comment	Organism
A246K	the mutant shows a large decrease in activity and a roughly 19fold lower affinity for myristic acid than the wild type enzyme	Bacillus subtilis
A246S	the mutation decreases the catalytic activity, but does not affect affinity for myristic acid	Bacillus subtilis
A246V	the mutant shows slightly reduced activity and moderately reduced affinity for myristic acid	Bacillus subtilis
F250K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
I244K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
I247K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
L237K	the mutants shows specific activity similar to the wild type enzyme	Bacillus subtilis
L241K	the mutants shows specific activity similar to the wild type enzyme	Bacillus subtilis
L251K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
P243A	the mutant shows decreased specific activity compared to the wild type enzyme and gives an absorption spectrum that is not characteristic of a nitrogenous ligand-bound form of a ferric P450	Bacillus subtilis
P243H	inactive, the mutant gives an absorption spectrum characteristic of a nitrogenous ligand-bound form of a ferric P450	Bacillus subtilis
P243K	inactive, the mutant gives an absorption spectrum characteristic of a nitrogenous ligand-bound form of a ferric P450	Bacillus subtilis
P243S	the mutant shows decreased specific activity compared to the wild type enzyme and gives an absorption spectrum that is not characteristic of a nitrogenous ligand-bound form of a ferric P450	Bacillus subtilis
R242A	the mutant shows about a 5fold lower affinity than the wild type for myristic acid, if cumene hydroperoxide is used instead of H2o2, however, the Km value is not affected much by this substitution	Bacillus subtilis
R242K	the mutant shows a large decrease in activity for myristic acid and H2O2, if cumene hydroperoxide is used instead of H2o2, however, the Km value is not affected much by this substitution	Bacillus subtilis
S248K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
V245K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis
Y249K	the mutant shows decreased specific activity compared to the wild type enzyme	Bacillus subtilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.021	-	H2O2	wild type enzyme, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis
0.11	-	cumene hydroperoxide	mutant enzyme R242A, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis
0.2	-	cumene hydroperoxide	wild type enzyme, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis
0.21	-	H2O2	mutant enzyme R242K, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis
0.24	-	cumene hydroperoxide	mutant enzyme R242K, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis
4.4	-	H2O2	mutant enzyme R242A, using myristic acid as cosubstrate, in 0.1 M potassium phosphate buffer (pH 6.7), at 25°C	Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
myristic acid + H2O2	Bacillus subtilis	CYP152A1 attacks the beta-carbon as well as the alpha-carbon of myristic acid	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	O31440	-	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA agarose column chromatography, gel filtration	Bacillus subtilis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
myristic acid + cumene hydroperoxide	-	Bacillus subtilis	?	-	?
myristic acid + H2O2	-	Bacillus subtilis	?	-	?
myristic acid + H2O2	CYP152A1 attacks the beta-carbon as well as the alpha-carbon of myristic acid	Bacillus subtilis	?	-	?

Synonyms

Synonyms	Comment	Organism
CYP152A1	-	Bacillus subtilis
peroxygenase cytochrome P450	-	Bacillus subtilis

Cofactor

Cofactor	Comment	Organism	Structure
heme	-	Bacillus subtilis

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Release 2023.1 (January 2023)