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Information on EC 3.2.1.217 - exo-acting protein-alpha-N-acetylgalactosaminidase
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EC Tree
3.2.1.217 exo-acting protein-alpha-N-acetylgalactosaminidaseIUBMB Comments
The enzyme, which belongs to the glycosylhydrolase 31 (GH31) family, is an exo-acting alpha-N-acetylgalactosaminidase that acts on the innermost alpha-GalNAc residues at the core of O-glycans when no other saccharides are attached to it. Unlike EC 3.2.1.49, alpha-N-acetylgalactosaminidase, it is not able to act on blood group A antigen.
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
+
=
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a [protein]-N-acetyl-alpha-D-galactosalaminyl-(L-serine/L-threonine)
+
=
a [protein]-(L-serine/L-threonine)
+
Synonyms
exo-acting alphaGalNAcase, GH31 alpha-GalNAcase, GH31 alpha-N-acetylgalactosaminidase, GH31 alphaGalNAcase, Nag31, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
GH31 alpha-GalNAcase
Nag31
Nag31
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a [protein]-N-acetyl-alpha-D-galactosalaminyl-(L-serine/L-threonine) + H2O = a [protein]-(L-serine/L-threonine) + N-acetyl-D-galactosamine
-
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
[protein]-N-acetyl-alpha-D-galactosalaminyl-(L-serine/L-threonine) N-acetylgalactosaminohydrolase
The enzyme, which belongs to the glycosylhydrolase 31 (GH31) family, is an exo-acting alpha-N-acetylgalactosaminidase that acts on the innermost alpha-GalNAc residues at the core of O-glycans when no other saccharides are attached to it. Unlike EC 3.2.1.49, alpha-N-acetylgalactosaminidase, it is not able to act on blood group A antigen.
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl alpha-N-acetyl-D-galactosaminide + H2O
4-methylumbelliferone + N-acetyl-D-galactosamine
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
4-methylumbelliferyl alpha-N-acetyl-D-galactosaminide + H2O
4-methylumbelliferone + N-acetyl-D-galactosamine
-
-
-
-
?
4-methylumbelliferyl alpha-N-acetyl-D-galactosaminide + H2O
4-methylumbelliferone + N-acetyl-D-galactosamine
-
-
-
-
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
-
-
-
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
-
-
-
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
-
-
-
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
-
-
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
-
-
-
?
BODIPY-IL29a peptide + H2O
N-acetyl-D-galactosamine + QPQPTAGPV
-
sequence QPQPT-(alpha-GalNAc)-AGPV
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-
?
BODIPY-IL29a peptide + H2O
N-acetyl-D-galactosamine + QPQPTAGPV
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sequence QPQPT-(alpha-GalNAc)-AGPV
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-
?
additional information
?
-
-
no activity with 4-methylumbelliferyl alpha-D-galactoside, 4-methylumbelliferyl alpha-D-glucoside, 4-methylumbelliferyl alpha-D-mannoside, 4-nitrophenyl alpha-D-xyloside, and 4-methylumbelliferyl A-antigen type 2tetra peptide
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-
-
additional information
?
-
-
no activity with GlcNAc-beta-1,3-GalNAcalpha-pNP
-
-
-
additional information
?
-
-
GalNAcalpha-L-Ser is a poor substrate
-
-
-
additional information
?
-
no activity toward 4-nitrophenyl alpha-D-glucoside, 4-nitrophenyl alpha-D-galactoside, 4-nitrophenyl alpha-N-acetyl-D-glucosamide, 4-nitrophenyl alpha-mannoside, and 4-nitrophenyl alpha-D-xyloside
-
-
-
additional information
?
-
-
no activity with 4-methylumbelliferyl alpha-D-galactoside, 4-methylumbelliferyl alpha-D-glucoside, 4-methylumbelliferyl alpha-D-mannoside, 4-nitrophenyl alpha-D-xyloside, and 4-methylumbelliferyl A-antigen type 2tetra peptide
-
-
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0172
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D508N, at pH 6.0 and 37°C
-
0.0232
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455N, at pH 6.0 and 37°C
-
0.042
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465A, at pH 7.0 and 37°C
-
0.046
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465S, at pH 7.0 and 37°C
-
0.06
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386F, at pH 6.0 and 37°C
-
0.0734
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455A, at pH 6.0 and 37°C
-
0.0794
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme L492R, at pH 6.0 and 37°C
-
0.101
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme V456A, at pH 6.0 and 37°C
-
0.11
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
0.136
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
truncated enzyme containing an FN3 domain but not C-terminal domains including a CBM32 domain, at pH 6.0 and 50°C
-
0.14
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
0.14
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
0.158
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type enzyme, at pH 6.0 and 37°C
-
0.158
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
full-length enzyme, at pH 6.0 and 50°C
-
0.27
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
0.34
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465G, at pH 7.0 and 37°C
-
2.23
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme I542F, at pH 6.0 and 37°C
-
2.36
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386A, at pH 6.0 and 37°C
-
3.86
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W221N, at pH 6.0 and 37°C
-
4.44
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W570A, at pH 6.0 and 37°C
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00231
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D508N, at pH 6.0 and 37°C
-
0.00239
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455N, at pH 6.0 and 37°C
-
0.01
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455A, at pH 6.0 and 37°C
-
0.0175
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme L492R, at pH 6.0 and 37°C
-
0.0506
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386A, at pH 6.0 and 37°C
-
0.0851
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386F, at pH 6.0 and 37°C
-
1.4
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme V456A, at pH 6.0 and 37°C
-
2.04
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme I542F, at pH 6.0 and 37°C
-
2.1
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
2.22
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
2.89
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W570A, at pH 6.0 and 37°C
-
3.6
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
3.79
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W221N, at pH 6.0 and 37°C
-
4.3
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465S, at pH 7.0 and 37°C
-
4.45
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
truncated enzyme containing an FN3 domain but not C-terminal domains including a CBM32 domain, at pH 6.0 and 50°C
-
5
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465A, at pH 7.0 and 37°C
-
5.7
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465G, at pH 7.0 and 37°C
-
6.54
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type enzyme, at pH 6.0 and 37°C
-
6.54
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
full-length enzyme, at pH 6.0 and 50°C
-
7.2
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.017
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465G, at pH 7.0 and 37°C
-
0.021
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386A, at pH 6.0 and 37°C
-
0.09
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465S, at pH 7.0 and 37°C
-
0.1
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455N, at pH 6.0 and 37°C
-
0.12
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
truncated mutant enzyme D465A, at pH 7.0 and 37°C
-
0.13
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D508N, at pH 6.0 and 37°C
-
0.14
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme D455A, at pH 6.0 and 37°C
-
0.22
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme L492R, at pH 6.0 and 37°C
-
0.65
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W570A, at pH 6.0 and 37°C
-
0.92
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme I542F, at pH 6.0 and 37°C
-
1
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme W221N, at pH 6.0 and 37°C
-
1.4
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme Y386F, at pH 6.0 and 37°C
-
14
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
mutant enzyme V456A, at pH 6.0 and 37°C
-
16
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
16
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type truncated enzyme, at pH 7.0 and 37°C
-
27
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
32
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type full-length enzyme, at pH 7.0 and 37°C
-
33
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
truncated enzyme containing an FN3 domain but not C-terminal domains including a CBM32 domain, at pH 6.0 and 50°C
-
41
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
-
wild type enzyme, at pH 6.0 and 37°C
-
41
4-nitrophenyl alpha-N-acetyl-D-galactosaminide
full-length enzyme, at pH 6.0 and 50°C
-
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 6.5
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). H7C6Z9_ENTFA
Enterococcus faecalis (strain ATCC 700802 / V583)
688
0
79596
TrEMBL
-
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
apoenzyme and in complex with alpha-N-acetyl-D-galactosaminide, hanging drop vapor diffusion method, using 25% (w/v) polyethylene glycol 3350 and 200 mM ammonium citrate tribasic
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D455A
-
the mutant shows 0.34% activity compared to the wild type enzyme
D455N
-
the mutant shows 0.24% activity compared to the wild type enzyme
D508N
-
the mutant shows 0.32% activity compared to the wild type enzyme
I542F
-
the mutant shows 2.2% activity compared to the wild type enzyme
L492R
-
the mutant shows 0.54% activity compared to the wild type enzyme
W221N
-
the mutant shows 2.4% activity compared to the wild type enzyme
W570A
-
the mutant shows 1.6% activity compared to the wild type enzyme
Y386A
-
the mutant shows 0.051% activity compared to the wild type enzyme
Y386F
-
the mutant shows 3.4% activity compared to the wild type enzyme
D465A
-
the mutant shows severely reduced activity compared to the wild type enzyme
D465G
-
the mutant shows severely reduced activity compared to the wild type enzyme
D465S
-
the mutant shows severely reduced activity compared to the wild type enzyme
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4 - 10
-
the enzyme is stable (more than 90% residual activity) at pH values ranging from 4.0 to 10.0 at 4°C for 20 h
764845
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography and Superdex 200 gel filtration
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
full-length enzyme and its truncated mutants are expressed in Escherichia coli BL21(DE3) cells
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the transcription of the enzyme increases significantly during the prepupal to pupal stage
-
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Miyazaki, T.; Ikegaya, M.; Alonso-Gil, S.
Structural and mechanistic insights into the substrate specificity and hydrolysis of GH31 alpha-N-acetylgalactosaminidase
Biochimie
195
90-99
2022
Enterococcus faecalis
brenda
Miyazaki, T.; Park, E.
Crystal structure of the Enterococcusxa0faecalis alpha-N-acetylgalactosaminidase, a member of the glycoside hydrolase family 31
FEBS Lett.
594
2282-2293
2020
Enterococcus faecalis (H7C6Z9)
brenda
Ikegaya, M.; Miyazaki, T.; Park, E.
Biochemical characterization of Bombyx mori alpha-N-acetylgalactosaminidase belonging to the glycoside hydrolase family 31
Insect Mol. Biol.
30
367-378
2021
Bombyx mori
brenda
Rahfeld, P.; Wardman, J.; Mehr, K.; Huff, D.; Morgan-Lang, C.; Chen, H.; Hallam, S.; Withers, S.
Prospecting for microbial alpha-N-acetylgalactosaminidases yields a new class of GH31 O-glycanase
J. Biol. Chem.
294
16400-16415
2019
Phocaeicola plebeius, Bacteroides caccae
brenda
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Release 2023.1 (January 2023)
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