Information on EC 1.14.13.195 - L-ornithine N5-monooxygenase (NADPH)

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
1.14.13.195
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RECOMMENDED NAME
GeneOntology No.
L-ornithine N5-monooxygenase (NADPH)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-ornithine + NADPH + H+ + O2 = N5-hydroxy-L-ornithine + NADP+ + H2O
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ferrichrome A biosynthesis
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pyoverdine I biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
L-ornithine,NADPH:oxygen oxidoreductase (N5-hydroxylating)
A flavoprotein (FAD). The enzyme is involved in biosynthesis of N5-hydroxy-L-ornithine, N5-formyl-N5-hydroxy-L-ornithine or N5-acetyl-N5-hydroxy-L-ornithine. These nonproteinogenic amino acids are building blocks of siderophores produced by some bacteria (e.g. Streptomyces coelicolor, Saccharopolyspora erythraea and Pseudomonas aeruginosa). The enzyme is specific for NADPH. cf. EC 1.14.13.196, L-ornithine N5-monooxygenase [NAD(P)H].
CAS REGISTRY NUMBER
COMMENTARY hide
93229-65-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene pvdA
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Manually annotated by BRENDA team
strains B10 and PF, gene pvdA
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Manually annotated by BRENDA team
strain KT2440, no activity in strain WCS358, gene pvdA
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Manually annotated by BRENDA team
strain B10, gene psbA
UniProt
Manually annotated by BRENDA team
strain B10, gene psbA
UniProt
Manually annotated by BRENDA team
pv. syringae, strain B278a, gene pvdA
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Manually annotated by BRENDA team
strain GMI1000, gene pvdA
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Manually annotated by BRENDA team
strain A3(2), gene cchB
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Manually annotated by BRENDA team
strain A3(2), gene cchB
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-lysine + NADPH + H+ + O2
L-lysine N6-oxide + NADP+ + H2O
show the reaction diagram
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-
-
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?
L-ornithine + NADH + H+ + O2
N5-hydroxy-L-ornithine + NAD+ + H2O
show the reaction diagram
Q5SE95
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-
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?
L-ornithine + NADPH + H+ + O2
L-ornithine N5-oxide + NADP+ + H2O
show the reaction diagram
L-ornithine + NADPH + H+ + O2
N5-hydroxy-L-ornithine + NADP+ + H2O
show the reaction diagram
L-ornithine + NADPH + H+ + O2
N5-hydroxy-L-ornithine + NADPH + H2O
show the reaction diagram
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
L-lysine + NADPH + H+ + O2
L-lysine N6-oxide + NADP+ + H2O
show the reaction diagram
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-
-
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?
L-ornithine + NADPH + H+ + O2
L-ornithine N5-oxide + NADP+ + H2O
show the reaction diagram
L-ornithine + NADPH + H+ + O2
N5-hydroxy-L-ornithine + NADP+ + H2O
show the reaction diagram
L-ornithine + NADPH + H+ + O2
N5-hydroxy-L-ornithine + NADPH + H2O
show the reaction diagram
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hydroxylation of the primary amine of ornithine in the initial step of the biosynthesis of siderophore pyoverdin
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?
additional information
?
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PvdA is the ornithine hydroxylase, which performs the first enzymatic step in preparation of hydroxamate siderophore derivatives, pyoverdin is the hydroxamate siderophore produced by the opportunistic pathogen Pseudomonas aeruginosa under the iron-limiting conditions of the human host, overview. Poor activity with DL-2,3-diaminopropionic acid and D-ornithine, no activity with L-norleucine, 5-aminopentanoic acid, DL-2,4-diaminobutyric acid, and 1,4-diaminobutane, and no activity with NADH as cofactor, substrate specificity, overview
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavin
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FAD or FMN, required, the reduction of FAD is ornithine-independent
additional information
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
the psbA expression is negatively controlled by iron
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chloromercuribenzoate
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a mixed inhibitor with respect to ornithine
5-aminopentanoic acid
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competitive inhibition
chloride
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mixed-type inhibitor with respect to ornithine, but a competitive inhibitor with respect to NADPH
L-2,4-diaminobutyrate
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competitive inhibition
L-homoserine
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competitive inhibition
L-lysine
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mixed-type inhibition
NADP+
competitive inhibitor
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
L-arginine
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enhances the rate of O2 activation, allosteric regulator
L-lysine
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nonsubstrate activator stimulating NADPh oxidation 3.9fold at 2 mM causing H2O2 formation, overview
L-ornithine
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activates 5fold
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0219
FAD
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pH 8.0, 25C, recombinant detagged enzyme
0.058 - 0.286
L-ornithine
0.1
NADH
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pH 7.5, 15C
0.012 - 0.8
NADPH
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.327
L-ornithine
Saccharopolyspora erythraea
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pH 8.0, 25C
0.73
NADH
Aspergillus fumigatus
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pH 7.5, 15C
0.12 - 0.85
NADPH
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.14
L-ornithine
Saccharopolyspora erythraea
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pH 8.0, 25C
192
10
NADH
Aspergillus fumigatus
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pH 7.5, 15C
8
0.34 - 50
NADPH
5
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9
5-aminopentanoic acid
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pH 8.0, 25C, recombinant detagged enzyme
8.4
L-2,4-diaminobutyrate
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pH 8.0, 25C, recombinant detagged enzyme
3.1
L-homoserine
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pH 8.0, 25C, recombinant detagged enzyme
additional information
additional information
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inhibition kinetics
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.24
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NADPH oxidase activity
0.321
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L-ornithine hydroxylation
0.534
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NADPH oxidation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
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pH profile, overview
6 - 9.5
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activity range, pH profile
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22 - 24
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assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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the enzyme anchors in the inner membrane with its N-terminus, while the bulk of the enzyme is located in the cytosol, the transmembrane region overlaps the FAd binding site, membrane-association determinants of PvdA and analysis of membrane topology, overview
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Manually annotated by BRENDA team
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a substantial amount of the enzyme is located in the membrane fraction. It is mainly clustered at the old cell pole of bacteria, indicating a polar segregation of the protein
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
136600
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recombinant His-tagged enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer or pentamer
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x * 51000, recombinant His-tagged enzyme, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
flavoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of the enzyme with FAD in oxidized (1.9 A resolution) and reduced (3.03 A resolution) states. Crystals are obtained through the hanging drop vapor diffusion method
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography, the tag is cleaved off by thrombin
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recombinant N-terminally His-tagged full-length and truncated wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography
recombinant N-terminally thioredoxin-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression as His6-tagged protein in Escherichia coli
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expression of PvdA-PhoA fusion protein mutants in Escherichia coli strain LMG194
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gene cchB, overexpression of N-terminally thioredoxin-tagged enzyme in Escherichia coli strain BL21(DE3)
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gene psbA, DNA and amino acid sequence determination and analysis, monocistronic gene
gene pvdA, expression of His-tagged enzyme in Escherichia coli strain Rosetta2(DE3)
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PvdA, DNA and amino acid sequence determination and analysis, sequence comparisons
PvdA, DNA and amino acid sequence determination and analysis, sequence comparisons, promoter activity analysis during cell growth. Recombinant expression of N-terminally His-tagged full-length and truncated wild-type and mutant enzymes in Escherichia coli strain M15
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme expression is induced by iron
the psbA expression is negatively controlled by iron
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S257A
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the interaction between residue Ser257 and NADP(H) is essential for stabilization of the C4a-hydroperoxyflavin intermediate. Ser257 may function as a pivot point, allowing the nicotinamide of NADP+ to slide into position for stabilization of the C4a-hydroperoxyflavin
additional information