Information on EC 1.14.14.40 - phenylalanine N-monooxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.14.40
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RECOMMENDED NAME
GeneOntology No.
phenylalanine N-monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-phenylalanine + 2 [reduced NADPH-hemoprotein reductase] + 2 O2 = (E)-phenylacetaldoxime + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
show the reaction diagram
overall reaction
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L-phenylalanine + [reduced NADPH—hemoprotein reductase] + O2 = N-hydroxy-L-phenylalanine + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
(1a)
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N,N-dihydroxy-L-phenylalanine = (E)-phenylacetaldoxime + CO2 + H2O
show the reaction diagram
(1c)
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N-hydroxy-L-phenylalanine + [reduced NADPH-hemoprotein reductase] + O2 = N,N-dihydroxy-L-phenylalanine + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
(1b)
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Cyanoamino acid metabolism
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Glucosinolate biosynthesis
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
L-phenylalanine,[reduced NADPH-hemoprotein reductase]:oxygen oxidoreductase (N-hydroxylating)
This cytochrome P-450 (heme-thiolate) enzyme, found in plants, catalyses two successive N-hydroxylations of L-phenylalanine, a committed step in the biosynthesis of benzylglucosinolate and the cyanogenic glucosides (R)-prunasin and (R)-amygdalin. The product of the two hydroxylations, N,N-dihydroxy-L-phenylalanine, is labile and undergoes dehydration followed by decarboxylation, producing an oxime. It is still not known whether the decarboxylation is spontaneous or catalysed by the enzyme.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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CYP79A2 is involved in the biosynthesis of benzylglucosinolate
physiological function
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enzyme is an N-hydroxylase converting L-phenylalanine into phenylacetaldoxime, the precursor of benzylglucosinolate. Transgenic Arabidopsis thaliana constitutively expressing CYP79A2 accumulate high levels of benzylglucosinolate
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-4-hydroxyphenylacetaldoxime + O2 + [reduced NADPH-hemoprotein reductase]
1-aci-nitro-2-(4-hydroxyphenyl)-ethane + H2O + [oxidized NADPH—hemoprotein reductase]
show the reaction diagram
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first step of reaction
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1-aci-nitro-2-(4-hydroxyphenyl)-ethane + 2-mercaptoethanol
(Z)-2-hydroxyethyl N-hydroxy-2-(4-hydroxyphenyl)ethanimidothioate + H2O
show the reaction diagram
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an (E)-omega-(methylthio)alkanal oxime + O2 + glutathione + [reduced NADPH-hemoprotein reductase]
an (E)-1-(glutathione-S-yl)-omega-(methylthio)alkylhydroximate + 2 H2O + [oxidized NADPH-hemoprotein reductase]
show the reaction diagram
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overall reaction
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?
L-phenylalanine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
(E)-phenylacetaldoxime + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
show the reaction diagram
L-phenylalanine + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
(E)-phenylacetaldoxime + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
show the reaction diagram
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?
tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
(E)-4-hydroxyphenylacetaldoxime + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
show the reaction diagram
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overall reaction, CYP79A1 catalyzes the specific production of (E)-phydroxyphenylacetaldoxime
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(E)-4-hydroxyphenylacetaldoxime + O2 + [reduced NADPH-hemoprotein reductase]
1-aci-nitro-2-(4-hydroxyphenyl)-ethane + H2O + [oxidized NADPH—hemoprotein reductase]
show the reaction diagram
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first step of reaction
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?
1-aci-nitro-2-(4-hydroxyphenyl)-ethane + 2-mercaptoethanol
(Z)-2-hydroxyethyl N-hydroxy-2-(4-hydroxyphenyl)ethanimidothioate + H2O
show the reaction diagram
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?
an (E)-omega-(methylthio)alkanal oxime + O2 + glutathione + [reduced NADPH-hemoprotein reductase]
an (E)-1-(glutathione-S-yl)-omega-(methylthio)alkylhydroximate + 2 H2O + [oxidized NADPH-hemoprotein reductase]
show the reaction diagram
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overall reaction
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?
additional information
?
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CYP83B1 catalyzes the conversion of the (E)-p-hydroxyphenylacetaldoxime into an S-alkyl-thiohydroximate with retention of the configuration of the E-oxime intermediate in the final glucosinolate core structure. CYP83B1 from Arabidopsis thaliana cannot convert the (E)-p-hydroxyphenylacetaldoxime to the (Z)-isomer, which blocks the route towards cyanogenic glucoside synthesis
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glutathione
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reduced glutathione, final concentration of 3 mM, slight stimulation
reduced glutathione
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addition of reduced glutathione to a final concentration of 3 mM stimulates aldoxime production
Tween 80
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addition of Tween 80 to a final concentration of 0.083% (v/v) results in a 1.5fold increase in aldoxime production; final concentration of 0.083% (v/v), 1.5fold increase in aldoxime production
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0067
L-phenylalanine
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pH 7.5, 26°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.9
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compared with the activity at pH 7.5, the activity of chimeric CYP79A2 is 25% at pH 6.0, 50% at pH 6.5, 80% at pH 7.0, and 70% at pH 7.9
6
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25% of maximum activity
7.9
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70% of maximum activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
expressed specifically in the seedling, coexpression with CYP71AN24 which catalyzes the conversion of phenylacetaldoxime into mandelonitrile with broad substrate specificity
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 60000, SDS-PAGE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli JM109 cells; expression in Escherichia coli
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expression in Saccharomyces cerevisiae
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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biosynthetic pathway for the production of phenylacetonitrile in Escherichia coli utilizing enzymes from the plant glucosinolate-biosynthetic and bacterial aldoxime-nitrile pathways, i.e. introducing the genes encoding cytochrome P450 (CYP) 79A2 and CYP reductase from Arabidopsis thaliana, yielding the E,Z-phenylacetaldoxime-producing transformant and further introducing the aldoxime dehydratase (Oxd) gene from Bacillus sp. strain OxB-1, yielding the phenylacetonitrile-producing transformant. Expression of active CYP79A2 and concentration of biomass are improved by the combination of the autoinduction method, coexpression of groE, encoding the heat shock protein GroEL/GroES, N-terminal truncation of CYP79A2, and optimization of the culture conditions, yielding a more than 60fold concentration of E,Z-phenylacetaldoxime up to 2.9 mM