ID   PALH_ERWRD              Reviewed;         453 AA.
AC   Q9AI65;
DT   24-NOV-2009, integrated into UniProtKB/Swiss-Prot.
DT   24-NOV-2009, sequence version 2.
DT   02-SEP-2026, entry version 81.
DE   RecName: Full=Alpha-glucosidase;
DE            EC=3.2.1.20;
GN   Name=palH;
OS   Erwinia rhapontici (Pectobacterium rhapontici).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Enterobacterales; Erwiniaceae; Erwinia.
OX   NCBI_TaxID=55212;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION IN PALATINOSE METABOLISM,
RP   PATHWAY, AND INDUCTION.
RC   STRAIN=ATCC 29283 / DSM 4484 / LMG 2688 / NCPPB 1578 / ICPB ER102 /
RC   CP/28;
RX   PubMed=11274100; DOI=10.1128/jb.183.8.2425-2430.2001;
RA   Boernke F., Hajirezaei M., Sonnewald U.;
RT   "Cloning and characterization of the gene cluster for palatinose metabolism
RT   from the phytopathogenic bacterium Erwinia rhapontici.";
RL   J. Bacteriol. 183:2425-2430(2001).
RN   [2]
RP   PROTEIN SEQUENCE OF 2-26, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY,
RP   COFACTOR, ACTIVITY REGULATION, KINETIC PARAMETERS, MASS SPECTROMETRY,
RP   SUBUNIT, AND MUTAGENESIS OF GLU-173 AND ILE-174.
RC   STRAIN=ATCC 29283 / DSM 4484 / LMG 2688 / NCPPB 1578 / ICPB ER102 /
RC   CP/28;
RX   PubMed=19625389; DOI=10.1093/molbev/msp162;
RA   Hall B.G., Pikis A., Thompson J.;
RT   "Evolution and biochemistry of family 4 glycosidases: implications for
RT   assigning enzyme function in sequence annotations.";
RL   Mol. Biol. Evol. 26:2487-2497(2009).
CC   -!- FUNCTION: Alpha-glucosidase with broad specificity. Hydrolyzes maltose,
CC       palatinose, maltulose, trehalose, trehalulose, turanose, leucrose,
CC       sucrose and maltitol. Is not active against alpha-galactosides, e.g.
CC       melibiose, and alpha-mannosides. Shows an obligate requirement for an
CC       O-alpha-glycosidic linkage, since it is not able to cleave beta-
CC       glycosidic bonds (cellobiose, gentiobiose, lactose, sophorose or
CC       laminaribiose). Cannot hydrolyze phosphorylated alpha-glucosides
CC       derivatives. Seems to be involved in the degradation of palatinose, a
CC       sucrose isomer that is formed as a reserve material under conditions of
CC       excess carbon availability, sequestered in a form unavailable to
CC       competitors such as fungi or the host plant, and whose consumption
CC       appears to be postponed until the preferentially metabolized carbon
CC       source (e.g. sucrose) is depleted. {ECO:0000269|PubMed:11274100}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Hydrolysis of terminal, non-reducing (1->4)-linked alpha-D-
CC         glucose residues with release of alpha-D-glucose.; EC=3.2.1.20;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC   -!- COFACTOR:
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Sr(2+); Xref=ChEBI:CHEBI:35104;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Note=Binds 1 divalent metal ion per subunit. Mn(2+) is the most
CC       efficient metal, but to a lesser extent, can also use Co(2+), Ca(2+),
CC       Fe(2+), Mg(2+), Sr(2+), and Ni(2+). Cannot use Zn(2+), Cu(2+) or
CC       Cr(2+). {ECO:0000269|PubMed:19625389};
CC   -!- COFACTOR:
CC       Name=NAD(+); Xref=ChEBI:CHEBI:57540;
CC         Evidence={ECO:0000269|PubMed:19625389};
CC       Note=Binds 1 NAD(+) per subunit. {ECO:0000269|PubMed:19625389};
CC   -!- ACTIVITY REGULATION: Is inhibited by EDTA in vitro.
CC       {ECO:0000269|PubMed:19625389}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=8.58 mM for maltose {ECO:0000269|PubMed:19625389};
CC         KM=7.31 mM for maltitol {ECO:0000269|PubMed:19625389};
CC         KM=6.59 mM for trehalose {ECO:0000269|PubMed:19625389};
CC         KM=3.63 mM for trehalulose {ECO:0000269|PubMed:19625389};
CC         KM=22.11 mM for sucrose {ECO:0000269|PubMed:19625389};
CC         KM=10.04 mM for turanose {ECO:0000269|PubMed:19625389};
CC         KM=1.75 mM for maltulose {ECO:0000269|PubMed:19625389};
CC         KM=5.31 mM for leucrose {ECO:0000269|PubMed:19625389};
CC         KM=2.52 mM for palatinose {ECO:0000269|PubMed:19625389};
CC         KM=180 uM for 4-nitrophenyl-alpha-D-glucopyranoside
CC         {ECO:0000269|PubMed:19625389};
CC         KM=104 uM for NAD(+) {ECO:0000269|PubMed:19625389};
CC         KM=47.2 uM for Mn(2+) {ECO:0000269|PubMed:19625389};
CC         Vmax=1.01 umol/min/mg enzyme with maltose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.24 umol/min/mg enzyme with maltitol as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.51 umol/min/mg enzyme with trehalose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.47 umol/min/mg enzyme with trehalulose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.31 umol/min/mg enzyme with sucrose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.46 umol/min/mg enzyme with turanose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.53 umol/min/mg enzyme with maltulose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.44 umol/min/mg enzyme with leucrose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=0.7 umol/min/mg enzyme with palatinose as substrate
CC         {ECO:0000269|PubMed:19625389};
CC         Vmax=6.01 umol/min/mg enzyme with
CC         4-nitrophenyl-alpha-D-glucopyranoside as substrate
CC         {ECO:0000269|PubMed:19625389};
CC   -!- PATHWAY: Glycan degradation; palatinose degradation.
CC       {ECO:0000269|PubMed:11274100}.
CC   -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:19625389}.
CC   -!- INDUCTION: Down-regulated by sucrose and up-regulated by palatinose.
CC       {ECO:0000269|PubMed:11274100}.
CC   -!- MASS SPECTROMETRY: Mass=50216; Method=Electrospray;
CC       Evidence={ECO:0000269|PubMed:19625389};
CC   -!- SIMILARITY: Belongs to the glycosyl hydrolase 4 family. {ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=AAK28734.1; Type=Erroneous initiation; Evidence={ECO:0000305};
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DR   EMBL; AF279280; AAK28734.1; ALT_INIT; Genomic_DNA.
DR   RefSeq; WP_133841662.1; NZ_AP024331.1.
DR   AlphaFoldDB; Q9AI65; -.
DR   SMR; Q9AI65; -.
DR   CAZy; GH4; Glycoside Hydrolase Family 4.
DR   OrthoDB; 9767022at2; -.
DR   BRENDA; 3.2.1.20; 2149.
DR   BRENDA; 3.2.1.86; 2149.
DR   SABIO-RK; Q9AI65; -.
DR   UniPathway; UPA01004; -.
DR   GO; GO:0004558; F:alpha-1,4-glucosidase activity; IEA:UniProtKB-EC.
DR   GO; GO:0016616; F:oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor; IEA:InterPro.
DR   GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR   Gene3D; 3.90.1820.10; AglA-like glucosidase; 1.
DR   InterPro; IPR053715; GH4_Enzyme_sf.
DR   InterPro; IPR001088; Glyco_hydro_4.
DR   InterPro; IPR022616; Glyco_hydro_4_C.
DR   InterPro; IPR015955; Lactate_DH/Glyco_Ohase_4_C.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   PANTHER; PTHR32092; 6-PHOSPHO-BETA-GLUCOSIDASE-RELATED; 1.
DR   PANTHER; PTHR32092:SF6; ALPHA-GALACTOSIDASE; 1.
DR   Pfam; PF02056; Glyco_hydro_4; 1.
DR   Pfam; PF11975; Glyco_hydro_4C; 1.
DR   PRINTS; PR00732; GLHYDRLASE4.
DR   SUPFAM; SSF56327; LDH C-terminal domain-like; 1.
DR   SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
PE   1: Evidence at protein level;
KW   Calcium; Carbohydrate metabolism; Cobalt; Direct protein sequencing;
KW   Glycosidase; Hydrolase; Iron; Magnesium; Manganese; Metal-binding; NAD;
KW   Nickel.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:19625389"
FT   CHAIN           2..453
FT                   /note="Alpha-glucosidase"
FT                   /id="PRO_0000389548"
FT   ACT_SITE        172
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000250"
FT   BINDING         3..69
FT                   /ligand="NAD(+)"
FT                   /ligand_id="ChEBI:CHEBI:57540"
FT                   /evidence="ECO:0000250"
FT   BINDING         149
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         171
FT                   /ligand="Mn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29035"
FT                   /evidence="ECO:0000250"
FT   BINDING         201
FT                   /ligand="Mn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29035"
FT                   /evidence="ECO:0000250"
FT   MUTAGEN         173
FT                   /note="E->S: Loss of catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:19625389"
FT   MUTAGEN         174
FT                   /note="I->V: No effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:19625389"
SQ   SEQUENCE   453 AA;  50341 MW;  64B178085AE436AC CRC64;
     MATKIVLVGA GSAQFGYGTL GDIFQSRALY GSEIILHDIN PVALAVTEKT AKDFLAKEDL
     PFIVSATTDR RTALRGAEFV IISIEVGDRF ALWDLDWQIP QQYGIQQVYG ENGGPGGLFH
     SLRIIPPILD ICADVADICP DAWIFNYSNP MSRICTTVHR RFPELNFVGM CHEIASLERY
     LPEMLNTSFD NLSLRAGGLN HFSVLLDARY KDSGKDAYAD VRAKAPDYFA SLPGYSDILA
     YTRQHGKLVD TEGSTERHAL GGKDSSYPWA DRTLFKEILE KFHCMPITVD SHFGEYISWA
     GEVSDHRGIL DFYTFYRNYL GGVQPKIELK LKERVVSIME GILTDSGYEE AAVNIPNRGF
     IKQLPEFIAV EVPAIIDRKG VHGIQVDIPP GIGGLLSNQI AIHDLTAEAI IAGSRDLVIQ
     ALLVDSVNNQ CRAIPELVDV MISRQQPWLN YLK
//