Any feedback?
No. of entries
Information on EC 3.2.1.217 - exo-acting protein-alpha-N-acetylgalactosaminidase
for references in articles please use BRENDA:EC3.2.1.217
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree 3 Hydrolases
3.2 Glycosylases
3.2.1 Glycosidases, i.e. enzymes that hydrolyse O- and S-glycosyl compounds
3.2.1.217 exo-acting protein-alpha-N-acetylgalactosaminidase
IUBMB Comments
The enzyme, which belongs to the glycosylhydrolase 31 (GH31) family, is an exo-acting α-N-acetylgalactosaminidase that acts on the innermost α-GalNAc residues at the core of O-glycans when no other saccharides are attached to it. Unlike EC 3.2.1.49, α-N-acetylgalactosaminidase, it is not able to act on blood group A antigen.
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
show+
=
+
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, more
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
GH31 alpha-GalNAcase
Nag31
Nag31
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
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
fetuin + H2O
N-acetyl-D-galactosamine + ?
-
Substrates: -
Products: -
Products: -
?
Tn antigen + H2O
N-acetyl-D-galactosamine + ?
-
Substrates: low activity
Products: -
Products: -
?
4-methylumbelliferyl alpha-N-acetyl-D-galactosaminide + H2O
4-methylumbelliferone + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
4-methylumbelliferyl alpha-N-acetyl-D-galactosaminide + H2O
4-methylumbelliferone + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
Substrates: -
Products: -
Products: -
?
4-nitrophenyl alpha-N-acetyl-D-galactosaminide + H2O
4-nitrophenol + N-acetyl-D-galactosamine
-
Substrates: -
Products: -
Products: -
?
BODIPY-IL29a peptide + H2O
N-acetyl-D-galactosamine + QPQPTAGPV
-
Substrates: sequence QPQPT-(alpha-GalNAc)-AGPV
Products: -
Products: -
?
BODIPY-IL29a peptide + H2O
N-acetyl-D-galactosamine + QPQPTAGPV
-
Substrates: sequence QPQPT-(alpha-GalNAc)-AGPV
Products: -
Products: -
?
additional information
?
-
-
Substrates: 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
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with GlcNAc-beta-1,3-GalNAcalpha-pNP
Products: -
Products: -
?
additional information
?
-
-
Substrates: GalNAcalpha-L-Ser is a poor substrate
Products: -
Products: -
?
additional information
?
-
Substrates: 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
Products: -
Products: -
?
additional information
?
-
-
Substrates: 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
Products: -
Products: -
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 6.5
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
other Location (Reliability: 4)
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography and Superdex 200 gel filtration
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the transcription of the enzyme increases significantly during the prepupal to pupal stage
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
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
Bacteroides caccae, Phocaeicola plebeius
brenda
EXTERNAL LINKS (specific for EC-Number 3.2.1.217)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2026.1 (March 2026)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
![DSMZ Digital Diversity]()
![Global Core Biodata Resource]()
![ELIXIR Core Data Resource]()
![German Network for Bioinformatics Infrastructure]()
