Information on EC 3.1.1.58 - N-acetylgalactosaminoglycan deacetylase

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

EC NUMBER
COMMENTARY hide
3.1.1.58
-
RECOMMENDED NAME
GeneOntology No.
N-acetylgalactosaminoglycan deacetylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
N-acetyl-D-galactosaminoglycan + H2O = D-galactosaminoglycan + acetate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of carboxylic ester
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
N-acetyl-D-galactosaminoglycan acetylhydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
52410-59-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
AHU 7139
-
-
Manually annotated by BRENDA team
AHU 7139
-
-
Manually annotated by BRENDA team
AHU 7165
-
-
Manually annotated by BRENDA team
gene hfsH
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
gene hfsH
-
-
Manually annotated by BRENDA team
Filobasidiella neoformans
-
-
Manually annotated by BRENDA team
Phage Vi
-
-
-
Manually annotated by BRENDA team
gene icaB
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-carboxyumbelliferyl acetate + H2O
3-carboxyumbelliferol + acetate
show the reaction diagram
-
-
-
-
?
beta-1,6-linked N-acetylglucosamine hexamer + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine hexamer + acetate
show the reaction diagram
-
-
-
-
?
beta-1,6-linked N-acetylglucosamine oligomer + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine oligomer + acetate
show the reaction diagram
-
the enzyme preferentially de-N-acetylates the second residue from the reducing terminus in the pentasaccharide and second and third residues from the reducing terminus in the hexasaccharide
-
-
?
beta-1,6-linked N-acetylglucosamine oligomers + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine oligomer + acetate
show the reaction diagram
-
specific for the substrate
-
-
?
beta-1,6-linked N-acetylglucosamine pentamer + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine pentamer + acetate
show the reaction diagram
-
-
-
-
?
beta-1,6-linked N-acetylglucosamine trimer + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine trimer + acetate
show the reaction diagram
-
-
-
-
?
N-acetylated beta-(1->6)-N-acetylglucosamine polymer + H2O
N-deacetylated beta-(1->6)-N-acetylglucosamine polymer + acetate
show the reaction diagram
-
the enzyme produces partially de-N-acetylated beta-(1->6)-N-acetylglucosamine polymers
-
-
?
N-acetylated oligogalactosamine + H2O
?
show the reaction diagram
N-acetylgalactosaminoglycan + H2O
galactosaminoglycan + acetate
show the reaction diagram
poly-beta-1,6-N-acetyl-D-glucosamine-adhesin + H2O
acetate + ?
show the reaction diagram
additional information
?
-
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
beta-1,6-linked N-acetylglucosamine oligomers + H2O
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine oligomer + acetate
show the reaction diagram
-
specific for the substrate
-
-
?
N-acetylated beta-(1->6)-N-acetylglucosamine polymer + H2O
N-deacetylated beta-(1->6)-N-acetylglucosamine polymer + acetate
show the reaction diagram
-
the enzyme produces partially de-N-acetylated beta-(1->6)-N-acetylglucosamine polymers
-
-
?
N-acetylgalactosaminoglycan + H2O
galactosaminoglycan + acetate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
activates
molybdate
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
-
stimulates, optimal concentration: 80 mM, pH 9.3: inhibition
Co2+
-
pH 5.3: stimulates, optimal concentration: 80 mM, pH 9.3: inhibition
Cu2+
-
85% inhibition at 1 mM
dipicolinic acid
-
a metal chelator, complete inhibition
EDTA
-
a metal chelator, complete inhibition
methyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-deoxy-2-[[(octanoylsulfanyl)acetyl]amino]-beta-D-lucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranoside
-
-
methyl 2-deoxy-2-(sulfamoylamino)-beta-D-glucopyranoside
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9% inhibition at 1 mM
methyl 2-deoxy-2-[(hydroxyacetyl)(methyl)amino]-beta-D-glucopyranoside
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84% inhibition at 1 mM
methyl 2-deoxy-2-[(hydroxyacetyl)amino]-beta-D-glucopyranoside
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8% inhibition at 1 mM
methyl 2-deoxy-2-[(methylsulfonyl)amino]-beta-D-glucopyranoside
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7% inhibition at 1 mM
methyl 2-deoxy-2-[(sulfanylacetyl)amino]-beta-D-glucopyranoside
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methyl 2-deoxy-2-[methyl(methylsulfonyl)amino]-beta-D-glucopyranoside
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18% inhibition at 1 mM
methyl 2-deoxy-2-[methyl(sulfamoyl)amino]-beta-D-glucopyranoside
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67% inhibition at 1 mM
methyl 2-deoxy-2-[methyl(sulfanylacetyl)amino]-beta-D-glucopyranoside
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48% inhibition at 1 mM
Mg2+
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pH 5.3: stimulates, optimal concentration: 80 mM, pH 9.3: inhibition
Mn2+
-
pH 5.3: stimulates, optimal concentration: 80 mM, pH 9.3: inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ammonium acetate buffer
Phage Vi
-
less effective than Tris buffer
-
Tris buffer
Phage Vi
-
enzyme is most active in Tris buffer
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5
3-carboxyumbelliferyl acetate
-
recombinant enzyme, pH 7.5, 37°C
20
beta-1,6-linked N-acetylglucosamine pentamer
-
recombinant enzyme, pH 7.5, 37°C
30
beta-1,6-linked N-acetylglucosamine trimer
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recombinant enzyme, pH 7.5, 37°C
0.54
N-acetylated polygalactosamine
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30°C, pH 5.3 and pH 9.3
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0.0159 - 0.0161
N-acetylgalactosamine
additional information
additional information
-
steady-state kinetics, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0013
3-carboxyumbelliferyl acetate
-
recombinant enzyme, pH 7.5, 37°C
0.0007
beta-1,6-linked N-acetylglucosamine pentamer
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recombinant enzyme, pH 7.5, 37°C
0.0007
beta-1,6-linked N-acetylglucosamine trimer
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recombinant enzyme, pH 7.5, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0025
3-carboxyumbelliferyl acetate
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recombinant enzyme, pH 7.5, 37°C
0.000003
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine pentamer
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recombinant enzyme, pH 7.5, 37°C
0.00002
partially de-N-acetylated beta-1,6-linked N-acetylglucosamine trimer
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recombinant enzyme, pH 7.5, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.28
methyl 2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-deoxy-2-[[(octanoylsulfanyl)acetyl]amino]-beta-D-lucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl-(1->6)-2-(acetylamino)-2-deoxy-beta-D-glucopyranoside
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pH and temperature not specified in the publication
0.32
methyl 2-deoxy-2-[(hydroxyacetyl)(methyl)amino]-beta-D-glucopyranoside
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pH and temperature not specified in the publication
0.48
methyl 2-deoxy-2-[(sulfanylacetyl)amino]-beta-D-glucopyranoside
-
pH and temperature not specified in the publication
5.8
methyl 2-deoxy-2-[methyl(methylsulfonyl)amino]-beta-D-glucopyranoside
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pH and temperature not specified in the publication
0.68
methyl 2-deoxy-2-[methyl(sulfamoyl)amino]-beta-D-glucopyranoside
-
pH and temperature not specified in the publication
0.92
methyl 2-deoxy-2-[methyl(sulfanylacetyl)amino]-beta-D-glucopyranoside
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
and pH 9.0, 2 optima
5.3
-
and pH 9.3, 2 optima
7.8
Phage Vi
-
-
9
-
and pH 5.0, 2 optima
9.3
-
and pH 5.3, 2 optima
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
most of the enzyme is loosely bound, only 14% are strongly bound
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28174
-
x * 28174, calculated from the deduced amino acid sequence
33000
-
recombinant enzyme, gel filtration
33113
-
1 * 33113, recombinant His10-tagged enzyme, sequence calculation
76000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 28174, calculated from the deduced amino acid sequence
monomer
-
1 * 33113, recombinant His10-tagged enzyme, sequence calculation
additional information
-
the enzyme contains a conserved (alpha/beta)8 fold
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of 24-28 mg/ml protein in 25 mM Tris base, pH 8.0, 300 mM NaCl, and 10% glycerol with 0.002 ml reservoir solution containing 0.1 M Tris base, pH 8.0, 0.2 M NaCl, 30-34% PEG 3350, and equilibration against a 0.5 ml reservoir solution, 1-2 days, X-ray diffraction structure determination and analysis at 2.20 A resolution, energy-dispersive X-ray spectroscopy
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purified recombinant enzyme, hanging drop vapour diffusion, protein in 50 mM HEPES-NaOH, pH 6.8, 200 mM NaCl is mixed with 25-30% w/v PEG 3350, 100 mM Tris-HCl, pH 7.5-8.0, X-ray diffraction structure determination and analysis at 1.90 A resolution, molecular replacement procedure, method overview. Construction of putative models of the C-terminal domain of the protein using a multitude of homology modelling algorithms, and test for the presence of signal in molecular replacement calculations, slow-cooling torsion-angle simulated annealing
alignment with PDB database: hits to tetratricopeptide repeat domains of human nucleoporin O-linked N-acetylglucosamine transferase, yeast mitochondrial outer membrane translocon protein Tom70p, and human peroxisomal targeting signal-1 receptor PEX5, putative domains: porin domain for export of poly-beta-1,6-N-acetyl-D-glucosamine, and periplasmic domain for protein-protein interactions
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
increased temperature might improve poly-beta-1,6-N-acetyl-D-glucosamine export in absence of deacetylation by PgaB
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
proteolytic cleavage of DUF187 domain truncation mutants PgaB516 and PgaB410 during overexpression in Escherichia coli
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18°C, stable for several months
-
-20°C, stable for several months
-
4°C, purified recombinant detagged enzyme at about 5 mg/ml, in purification buffer containing 5% v/v glycerol, 10 days with no significant loss of activity
-
4°C, stable for one week
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
140fold, AHU 7165
-
homogeneity
-
recombinant enzyme from Escherichia coli strain DE3 pLysS by cation exchange chromatography and gel filtration
recombinant expression of wild-type and mutant enzymes from Escherichia coli
-
recombinant N-terminally His10-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage through Factor Xa protease, and ulrafiltration/dialysis
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of enzyme deficiency mutant in host strain
-
expression in Escherichia coli strain DE3 pLysS
from chromosomal DNA in pUC19 and subsequently in pCR2.1-TOPO for site directed mutagenesis and complementation experiments
fusion protein with glutathione S-transferase
fusion protein with green fluorescent protein and beta-galactosidase
-
gene hfsH, sequence comparisons
-
gene hfsH, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli
-
gene icaB, recombinant overexpression of N-terminally His10-tagged enzyme lacking the native N-terminal signal peptide in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D48A
-
site-directed mutagenesis of the catalytic residue abolishes the esterase activity of the enzyme, the mutant strain is deficient in holdfast anchoring to the cell body, and small holdfasts are shed into the medium. The mutant enzyme has a similar secondary structure compared to the wild-type enzyme based on circular dichroism measurement
D115A
decreased catalytic activity, 6.4% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin
H184A
mutant, deficient in biofilm formation
PgaB271
DUF187 domain truncation mutant, amino acids 1-271, deficient in biofilm formation and poly-beta-1,6-N-acetyl-D-glucosamine secretion, 4.1% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin
PgaB410
DUF187 domain truncation mutant, amino acids 1-410, proteolytic cleavage during overexpression
PgaB516
DUF187 domain truncation mutant, amino acids 1-516, deficient in biofilm formation and poly-beta-1,6-N-acetyl-D-glucosamine secretion, 5% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin, proteolytic cleavage during overexpression
D115A
-
decreased catalytic activity, 6.4% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin
-
H184A
-
mutant, deficient in biofilm formation
-
PgaB271
-
DUF187 domain truncation mutant, amino acids 1-271, deficient in biofilm formation and poly-beta-1,6-N-acetyl-D-glucosamine secretion, 4.1% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin
-
PgaB410
-
DUF187 domain truncation mutant, amino acids 1-410, proteolytic cleavage during overexpression
-
PgaB516
-
DUF187 domain truncation mutant, amino acids 1-516, deficient in biofilm formation and poly-beta-1,6-N-acetyl-D-glucosamine secretion, 5% of wild-type activity on N-deacetylation of poly-beta-1,6-N-acetyl-D-glucosamine-adhesin, proteolytic cleavage during overexpression
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
-
the enzyme is a good target for therapeutic intervention in biofilm related infections
medicine
-
the enzyme is a potential therapeutic target for antibiofilm therapies