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Information on EC 2.3.1.201 - UDP-2-acetamido-3-amino-2,3-dideoxy-glucuronate N-acetyltransferase and Organism(s) Pseudomonas aeruginosa and UniProt Accession G3XD01

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IUBMB Comments
This enzyme participates in the biosynthetic pathway for UDP-alpha-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-mannuronic acid), an important precursor of B-band lipopolysaccharide.
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This record set is specific for:
Pseudomonas aeruginosa
UNIPROT: G3XD01
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The taxonomic range for the selected organisms is: Pseudomonas aeruginosa
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
pgn_0002, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
UDP-2-acetamido-3-amino-2,3-dideoxy-glucuronate N-acetyltransferase
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UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid3-N-acetyltransferase
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UDP-2-acetamido-3-amino-2,3-dideoxy-glucuronate N-acetyltransferase
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WlbB
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
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-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate N-acetyltransferase
This enzyme participates in the biosynthetic pathway for UDP-alpha-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-mannuronic acid), an important precursor of B-band lipopolysaccharide.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
CoA + UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronate
show the reaction diagram
-
-
-
?
acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
CoA + UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronate
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
CoA + UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronate
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
the enzyme is involved in the biosynthesis of a rare di-N-acetylated sugar in the lipopolysaccharide 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.107
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
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in 50 mM HEPES (pH 8.0), 2.5 mM dithiothreitol, and 2 mM MgCl2, at 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
48.3
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
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in 50 mM HEPES (pH 8.0), 2.5 mM dithiothreitol, and 2 mM MgCl2, at 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
-
pI-value of above 8.0, isoelectric focusing
8.2
-
calculated from amino acid sequence
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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the enzyme is required for B-band O-antigen biosynthesis in Pseudomonas aeruginosa
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22700
x * 22700, predicted from amino acid sequence
22726
23000
-
3 * 23000, His-tagged enzyme, SDS-PAGE
61000
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His-tagged enzyme, gel filtration
68180
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MALDI-TOF mass spectrometry
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 22700, predicted from amino acid sequence
homotrimer
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K136A
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the mutation reduces the stabilizing effects of acetyl-CoA
K136R
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the mutation showed no discernible effect the stabilizing effects of acetyl-CoA on the purified mutant protein
additional information
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construction of a gene wbpD knockout strain, functional complementation by expression of WbpB from Bordetella pertussis, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
incubation of the enzyme with acetyl-CoA significantly enhances the stability of the protein and prevents precipitation over a course of 14 days
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
nickel-nitrilotriacetic acid-agarose column chromatography
Ni-NTA agarose column chromatography
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Ni-NTA agarose resin column chromatography, gel filtration
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recombinant His6-tagged WbpD from Escherichia coli strain BL21(DE3)
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli Tuner(DE3) cells
expressed as an N-terminally histidine-tagged fusion protein in Escherichia coli BL21(DE3) pLysS cells
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expressed in Escherichia coli BL21-CodonPlus(DE3) RIL cells
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gene wbpD, expression and subcloning of His6-tagged WbpD in Escherichia coli strain BL21(DE3)
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wenzel, C.Q.; Daniels, C.; Keates, R.A.; Brewer, D.; Lam, J.S.
Evidence that WbpD is an N-acetyltransferase belonging to the hexapeptide acyltransferase superfamily and an important protein for O-antigen biosynthesis in Pseudomonas aeruginosa PAO1
Mol. Microbiol.
57
1288-1303
2005
Pseudomonas aeruginosa
Manually annotated by BRENDA team
Westman, E.L.; Preston, A.; Field, R.A.; Lam, J.S.
Biosynthesis of a rare di-N-acetylated sugar in the lipopolysaccharides of both Pseudomonas aeruginosa and Bordetella pertussis occurs via an identical scheme despite different gene clusters
J. Bacteriol.
190
6060-6069
2008
Pseudomonas aeruginosa
Manually annotated by BRENDA team
Westman, E.L.; McNally, D.J.; Charchoglyan, A.; Brewer, D.; Field, R.A.; Lam, J.S.
Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa
J. Biol. Chem.
284
11854-11862
2009
Pseudomonas aeruginosa (G3XD01)
Manually annotated by BRENDA team
Larkin, A.; Imperiali, B.
Biosynthesis of UDP-GlcNAc(3NAc)A by WbpB, WbpE, and WbpD: enzymes in the Wbp pathway responsible for O-antigen assembly in Pseudomonas aeruginosa PAO1
Biochemistry
48
5446-5455
2009
Pseudomonas aeruginosa
Manually annotated by BRENDA team