BRENDA - Enzyme Database
show all sequences of 1.14.13.195

Heterologous expression, purification, and characterization of an L-ornithine N5-hydroxylase involved in pyoverdine siderophore biosynthesis in Pseudomonas aeruginosa

Ge, L.; Seah, S.Y.K.; J. Bacteriol. 188, 7205-7210 (2006)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
L-lysine
nonsubstrate activator stimulating NADPh oxidation 3.9fold at 2 mM causing H2O2 formation, overview
Pseudomonas aeruginosa
L-ornithine
activates 5fold
Pseudomonas aeruginosa
Cloned(Commentary)
Cloned (Commentary)
Organism
gene pvdA, expression of His-tagged enzyme in Escherichia coli strain Rosetta2(DE3)
Pseudomonas aeruginosa
Inhibitors
Inhibitors
Commentary
Organism
Structure
5-aminopentanoic acid
competitive inhibition
Pseudomonas aeruginosa
L-2,4-diaminobutyrate
competitive inhibition
Pseudomonas aeruginosa
L-homoserine
competitive inhibition
Pseudomonas aeruginosa
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten steady-state kinetic analysis, overview
Pseudomonas aeruginosa
0.0219
-
FAD
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
0.058
-
L-ornithine
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
0.16
-
NADPH
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
51000
-
x * 51000, recombinant His-tagged enzyme, SDS-PAGE
Pseudomonas aeruginosa
136600
-
recombinant His-tagged enzyme, gel filtration
Pseudomonas aeruginosa
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-ornithine + NADPH + H+ + O2
Pseudomonas aeruginosa
-
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas aeruginosa
-
gene pvdA
-
Posttranslational Modification
Posttranslational Modification
Commentary
Organism
flavoprotein
-
Pseudomonas aeruginosa
Purification (Commentary)
Purification (Commentary)
Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography, the tag is cleaved off by thrombin
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.24
-
NADPH oxidase activity
Pseudomonas aeruginosa
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
L-ornithine + NADPH + H+ + O2
-
704269
Pseudomonas aeruginosa
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
-
?
L-ornithine + NADPH + H+ + O2
no activity with D-ornithine
704269
Pseudomonas aeruginosa
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
-
?
additional information
the NADPH oxidase activity of the enzyme is tightly coupled to hydroxylamine formation
704269
Pseudomonas aeruginosa
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
tetramer or pentamer
x * 51000, recombinant His-tagged enzyme, SDS-PAGE
Pseudomonas aeruginosa
Synonyms
Synonyms
Commentary
Organism
L-ornithine N5-hydroxylase
-
Pseudomonas aeruginosa
PvdA
-
Pseudomonas aeruginosa
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Pseudomonas aeruginosa
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
-
Pseudomonas aeruginosa
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
9
pH profile, overview
Pseudomonas aeruginosa
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
specific for
Pseudomonas aeruginosa
additional information
no activity with FMN and NADH
Pseudomonas aeruginosa
NADPH
specific for
Pseudomonas aeruginosa
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
inhibition kinetics
Pseudomonas aeruginosa
2.9
-
5-aminopentanoic acid
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
3.1
-
L-homoserine
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
8.4
-
L-2,4-diaminobutyrate
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
L-lysine
nonsubstrate activator stimulating NADPh oxidation 3.9fold at 2 mM causing H2O2 formation, overview
Pseudomonas aeruginosa
L-ornithine
activates 5fold
Pseudomonas aeruginosa
Cloned(Commentary) (protein specific)
Commentary
Organism
gene pvdA, expression of His-tagged enzyme in Escherichia coli strain Rosetta2(DE3)
Pseudomonas aeruginosa
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
specific for
Pseudomonas aeruginosa
additional information
no activity with FMN and NADH
Pseudomonas aeruginosa
NADPH
specific for
Pseudomonas aeruginosa
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
5-aminopentanoic acid
competitive inhibition
Pseudomonas aeruginosa
L-2,4-diaminobutyrate
competitive inhibition
Pseudomonas aeruginosa
L-homoserine
competitive inhibition
Pseudomonas aeruginosa
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
inhibition kinetics
Pseudomonas aeruginosa
2.9
-
5-aminopentanoic acid
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
3.1
-
L-homoserine
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
8.4
-
L-2,4-diaminobutyrate
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten steady-state kinetic analysis, overview
Pseudomonas aeruginosa
0.0219
-
FAD
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
0.058
-
L-ornithine
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
0.16
-
NADPH
pH 8.0, 25°C, recombinant detagged enzyme
Pseudomonas aeruginosa
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
51000
-
x * 51000, recombinant His-tagged enzyme, SDS-PAGE
Pseudomonas aeruginosa
136600
-
recombinant His-tagged enzyme, gel filtration
Pseudomonas aeruginosa
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-ornithine + NADPH + H+ + O2
Pseudomonas aeruginosa
-
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
?
Posttranslational Modification (protein specific)
Posttranslational Modification
Commentary
Organism
flavoprotein
-
Pseudomonas aeruginosa
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain Rosetta2(DE3) by nickel affinity chromatography, the tag is cleaved off by thrombin
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.24
-
NADPH oxidase activity
Pseudomonas aeruginosa
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
L-ornithine + NADPH + H+ + O2
-
704269
Pseudomonas aeruginosa
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
-
?
L-ornithine + NADPH + H+ + O2
no activity with D-ornithine
704269
Pseudomonas aeruginosa
N5-hydroxy-L-ornithine + NADP+ + H2O
-
-
-
?
additional information
the NADPH oxidase activity of the enzyme is tightly coupled to hydroxylamine formation
704269
Pseudomonas aeruginosa
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer or pentamer
x * 51000, recombinant His-tagged enzyme, SDS-PAGE
Pseudomonas aeruginosa
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Pseudomonas aeruginosa
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
-
Pseudomonas aeruginosa
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
9
pH profile, overview
Pseudomonas aeruginosa
General Information
General Information
Commentary
Organism
physiological function
the enzyme is involved in pyoverdine siderophore biosynthesis, pyoverdine is required for acquiration of the essential iron from the host
Pseudomonas aeruginosa
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the enzyme is involved in pyoverdine siderophore biosynthesis, pyoverdine is required for acquiration of the essential iron from the host
Pseudomonas aeruginosa
Other publictions for EC 1.14.13.195
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
743137
Badieyan
Mechanism of N-hydroxylation ...
Aspergillus fumigatus, Aspergillus fumigatus Af293
J. Org. Chem.
80
2139-2147
2015
-
-
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-
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2
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2
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4
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2
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2
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4
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-
744163
Robinson
Contribution to catalysis of ...
Aspergillus fumigatus
Arch. Biochem. Biophys.
585
25-31
2015
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-
1
1
4
-
-
12
-
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1
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2
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1
1
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1
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2
1
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8
1
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1
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1
1
1
4
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12
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1
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1
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1
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1
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8
1
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1
1
-
8
8
725540
Shirey
Role of Ser-257 in the sliding ...
Aspergillus fumigatus
J. Biol. Chem.
288
32440-32448
2013
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1
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1
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4
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1
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4
-
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4
4
724451
Romero
Dual role of NADP(H) in the re ...
Aspergillus fumigatus
Biochim. Biophys. Acta
1824
850-857
2012
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-
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2
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2
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3
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2
-
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-
1
1
-
2
2
724860
Guillon
High cellular organization of ...
Pseudomonas aeruginosa
Environ. Microbiol.
14
1982-1994
2012
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1
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1
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1
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1
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714263
Robbel
Consecutive enzymatic modifica ...
Saccharopolyspora erythraea
Biochemistry
50
6073-6080
2011
-
-
1
-
-
-
-
1
-
-
-
1
-
1
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1
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2
-
1
1
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1
1
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2
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1
2
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1
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1
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1
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2
-
1
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-
1
1
-
-
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-
-
-
-
1
1
725195
Frederick
Regulated O2 activation in fla ...
Aspergillus fumigatus
J. Am. Chem. Soc.
133
12338-12341
2011
1
-
-
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2
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1
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1
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1
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1
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-
-
-
-
725440
Olucha
Two structures of an N-hydroxy ...
Pseudomonas aeruginosa
J. Biol. Chem.
286
31789-31798
2011
-
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1
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1
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3
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1
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1
1
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1
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1
1
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1
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1
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1
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1
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702303
Meneely
Kinetic mechanism of ornithine ...
Pseudomonas aeruginosa
Biochemistry
48
4371-4376
2009
-
-
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-
-
-
-
1
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1
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2
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1
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1
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3
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2
1
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1
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2
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2
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1
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1
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1
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-
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-
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3
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-
705555
Imperi
Membrane-association determina ...
Pseudomonas aeruginosa
Microbiology
154
2804-2813
2008
-
-
1
-
1
-
-
-
1
-
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4
-
1
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1
2
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2
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2
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1
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706031
Pohlmann
delta-Amino group hydroxylatio ...
Streptomyces coelicolor, Streptomyces coelicolor A3(2)
Org. Biomol. Chem.
6
1843-1848
2008
-
-
1
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1
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2
-
43
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1
1
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6
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1
1
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1
1
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2
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1
2
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1
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2
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1
1
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6
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1
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1
1
-
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1
1
-
-
-
702210
Meneely
Biochemical characterization o ...
Pseudomonas aeruginosa
Biochemistry
46
11930-11937
2007
-
-
-
-
-
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3
1
1
-
-
3
-
5
-
1
-
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-
2
-
5
1
2
-
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1
1
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3
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3
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3
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1
1
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3
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1
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2
-
5
1
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1
1
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-
-
-
-
704269
Ge
Heterologous expression, purif ...
Pseudomonas aeruginosa
J. Bacteriol.
188
7205-7210
2006
2
-
1
-
-
-
3
4
-
-
2
1
-
1
-
1
1
-
-
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1
-
3
1
2
1
-
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1
1
-
3
4
-
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2
-
1
3
-
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-
-
3
4
4
-
-
2
1
-
-
1
1
-
-
1
-
3
1
1
-
-
-
1
1
-
-
-
1
1
-
-
-
702009
Putignani
Expression of L-ornithine Ndel ...
Burkholderia cepacia, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae, Ralstonia solanacearum
Biochem. Biophys. Res. Commun.
313
245-257
2004
-
-
6
-
1
-
-
-
-
-
-
6
-
15
-
6
1
-
-
-
-
-
12
1
12
-
-
-
-
-
-
-
12
-
1
-
-
-
6
12
-
1
-
-
-
-
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-
-
-
6
-
-
6
1
-
-
-
-
12
1
-
-
-
-
-
-
-
1
5
-
-
5
-
-
704262
Ambrosi
Pseudobactin biogenesis, in th ...
Pseudomonas sp., Pseudomonas sp. B10
J. Bacteriol.
182
6233-6238
2000
-
-
1
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1
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1
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2
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2
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1
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2
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2
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1
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4
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1
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1
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