AGAL_BACTN
ID AGAL_BACTN Reviewed; 662 AA.
AC Q8A6L0;
DT 25-JAN-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2003, sequence version 1.
DT 03-AUG-2022, entry version 94.
DE RecName: Full=Retaining alpha-galactosidase;
DE EC=3.2.1.22;
DE AltName: Full=BtGH97b;
DE AltName: Full=Melibiase;
DE Flags: Precursor;
GN OrderedLocusNames=BT_1871;
OS Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / JCM 5827 /
OS CCUG 10774 / NCTC 10582 / VPI-5482 / E50).
OC Bacteria; Bacteroidetes; Bacteroidia; Bacteroidales; Bacteroidaceae;
OC Bacteroides.
OX NCBI_TaxID=226186;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 29148 / DSM 2079 / JCM 5827 / CCUG 10774 / NCTC 10582 /
RC VPI-5482 / E50;
RX PubMed=12663928; DOI=10.1126/science.1080029;
RA Xu J., Bjursell M.K., Himrod J., Deng S., Carmichael L.K., Chiang H.C.,
RA Hooper L.V., Gordon J.I.;
RT "A genomic view of the human-Bacteroides thetaiotaomicron symbiosis.";
RL Science 299:2074-2076(2003).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND KINETIC
RP PARAMETERS.
RC STRAIN=ATCC 29148 / DSM 2079 / JCM 5827 / CCUG 10774 / NCTC 10582 /
RC VPI-5482 / E50;
RX PubMed=18848471; DOI=10.1016/j.chembiol.2008.09.005;
RA Gloster T.M., Turkenburg J.P., Potts J.R., Henrissat B., Davies G.J.;
RT "Divergence of catalytic mechanism within a glycosidase family provides
RT insight into evolution of carbohydrate metabolism by human gut flora.";
RL Chem. Biol. 15:1058-1067(2008).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 27-662 IN COMPLEX WITH CALCIUM,
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP ACTIVITY REGULATION, ACTIVE SITE, MUTAGENESIS OF ASP-415 AND GLU-470,
RP CATALYTIC MECHANISM, AND SUBUNIT.
RC STRAIN=ATCC 29148 / DSM 2079 / JCM 5827 / CCUG 10774 / NCTC 10582 /
RC VPI-5482 / E50;
RX PubMed=19646996; DOI=10.1016/j.jmb.2009.07.068;
RA Okuyama M., Kitamura M., Hondoh H., Kang M.S., Mori H., Kimura A.,
RA Tanaka I., Yao M.;
RT "Catalytic mechanism of retaining alpha-galactosidase belonging to
RT glycoside hydrolase family 97.";
RL J. Mol. Biol. 392:1232-1241(2009).
CC -!- FUNCTION: Galactosidase that is able to hydrolyze the alpha-1,6
CC disaccharide melibiose and the synthetic p-nitrophenyl alpha-
CC galactoside substrate (pNP-Gal), with retention of the anomeric
CC configuration. Does not hydrolyze DNP-Glc or pNP-Glc.
CC {ECO:0000269|PubMed:18848471, ECO:0000269|PubMed:19646996}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal, non-reducing alpha-D-galactose
CC residues in alpha-D-galactosides, including galactose
CC oligosaccharides, galactomannans and galactolipids.; EC=3.2.1.22;
CC Evidence={ECO:0000269|PubMed:18848471, ECO:0000269|PubMed:19646996};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:19646996};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000269|PubMed:19646996};
CC -!- ACTIVITY REGULATION: Inhibited by EDTA in vitro.
CC {ECO:0000269|PubMed:19646996}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.31 mM for p-nitrophenyl alpha-galactoside (at pH 6.0 and 37
CC degrees Celsius) {ECO:0000269|PubMed:18848471,
CC ECO:0000269|PubMed:19646996};
CC KM=1.5 mM for melibiose (at pH 6.6 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:18848471, ECO:0000269|PubMed:19646996};
CC Note=kcat is 244 sec(-1) with p-nitrophenyl alpha-galactoside as
CC substrate (at pH 6.0 and 37 degrees Celsius).;
CC pH dependence:
CC Optimum pH is 8.0 with p-nitrophenyl alpha-galactoside as substrate.
CC {ECO:0000269|PubMed:19646996};
CC -!- SUBUNIT: Monomer. {ECO:0000305|PubMed:19646996}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 97 family. {ECO:0000305}.
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DR EMBL; AE015928; AAO76978.1; -; Genomic_DNA.
DR RefSeq; NP_810784.1; NC_004663.1.
DR RefSeq; WP_011108029.1; NC_004663.1.
DR PDB; 3A24; X-ray; 2.30 A; A/B=27-662.
DR PDB; 5E1Q; X-ray; 1.94 A; A/B=27-662.
DR PDBsum; 3A24; -.
DR PDBsum; 5E1Q; -.
DR AlphaFoldDB; Q8A6L0; -.
DR SMR; Q8A6L0; -.
DR STRING; 226186.BT_1871; -.
DR CAZy; GH97; Glycoside Hydrolase Family 97.
DR PaxDb; Q8A6L0; -.
DR PRIDE; Q8A6L0; -.
DR DNASU; 1075689; -.
DR EnsemblBacteria; AAO76978; AAO76978; BT_1871.
DR GeneID; 60927858; -.
DR KEGG; bth:BT_1871; -.
DR PATRIC; fig|226186.12.peg.1923; -.
DR eggNOG; COG4948; Bacteria.
DR HOGENOM; CLU_011166_0_0_10; -.
DR OMA; WLWAHWR; -.
DR BRENDA; 3.2.1.22; 709.
DR SABIO-RK; Q8A6L0; -.
DR EvolutionaryTrace; Q8A6L0; -.
DR Proteomes; UP000001414; Chromosome.
DR GO; GO:0030246; F:carbohydrate binding; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0052692; F:raffinose alpha-galactosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 2.60.40.1180; -; 1.
DR Gene3D; 2.70.98.10; -; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR014718; GH-type_carb-bd.
DR InterPro; IPR029483; GH97_C.
DR InterPro; IPR019563; GH97_catalytic.
DR InterPro; IPR029486; GH97_N.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR Pfam; PF14509; GH97_C; 1.
DR Pfam; PF14508; GH97_N; 1.
DR Pfam; PF10566; Glyco_hydro_97; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Glycosidase; Hydrolase;
KW Metal-binding; Reference proteome; Signal.
FT SIGNAL 1..19
FT /evidence="ECO:0000255"
FT CHAIN 20..662
FT /note="Retaining alpha-galactosidase"
FT /id="PRO_0000415272"
FT ACT_SITE 415
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:19646996"
FT ACT_SITE 470
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000269|PubMed:19646996"
FT BINDING 174
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:19646996"
FT BINDING 464
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:19646996"
FT BINDING 470
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:19646996"
FT MUTAGEN 415
FT /note="D->N,G: Loss of catalytic activity. The activity is
FT restored by adding an external nucleophilic azide ion."
FT /evidence="ECO:0000269|PubMed:19646996"
FT MUTAGEN 470
FT /note="E->Q: Loss of catalytic activity. No change in
FT substrate affinity."
FT /evidence="ECO:0000269|PubMed:19646996"
FT STRAND 30..48
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 51..63
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 70..72
FT /evidence="ECO:0007829|PDB:3A24"
FT STRAND 75..88
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 90..93
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 94..108
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 109..111
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 112..119
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 122..129
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 135..142
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 151..154
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 165..169
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 178..181
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 182..184
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 193..200
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 203..210
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 213..215
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 218..233
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 237..244
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 245..247
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 249..255
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 259..262
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 270..278
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 279..284
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 287..290
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 306..309
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 312..316
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 326..328
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 329..341
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 346..349
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 370..378
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 379..381
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 383..389
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 390..394
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 397..407
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 411..415
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 422..437
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 441..444
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 453..456
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 460..463
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 469..473
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 481..487
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 488..490
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 492..494
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 505..508
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 517..519
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 522..524
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 526..535
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 539..543
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 547..551
FT /evidence="ECO:0007829|PDB:5E1Q"
FT HELIX 554..562
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 568..576
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 577..579
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 580..587
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 590..597
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 602..607
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 616..623
FT /evidence="ECO:0007829|PDB:5E1Q"
FT TURN 625..629
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 634..641
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 646..651
FT /evidence="ECO:0007829|PDB:5E1Q"
FT STRAND 656..662
FT /evidence="ECO:0007829|PDB:5E1Q"
SQ SEQUENCE 662 AA; 74850 MW; 18A74C4AA328CE80 CRC64;
MKKLTFLLLC VLCTLSLQAQ KQFTLASPDG NLKTTITIGD RLTYDITCNG RQILTPSPIS
MTLDNGTVWG ENAKLSGTSR KSVDEMIPSP FYRASELRNH YNGLTLRFKK DWNVEFRAYN
DGIAYRFVNQ GKKPFRVVTE VSDYCFPSDM TASVPYVKSG KDGDYNSQFF NSFENTYTTD
KLSKLNKQRL MFLPLVVDAG DGVKVCITES DLENYPGLYL SASEGANRLS SMHAPYPKRT
VQGGHNQLQM LVKEHEDYIA KVDKPRNFPW RIAVVTTTDK DLAATNLSYL LGAPSRMSDL
SWIKPGKVAW DWWNDWNLDG VDFVTGVNNP TYKAYIDFAS ANGIEYVILD EGWAVNLQAD
LMQVVKEIDL KELVDYAASK NVGIILWAGY HAFERDMENV CRHYAEMGVK GFKVDFMDRD
DQEMTAFNYR AAEMCAKYKL ILDLHGTHKP AGLNRTYPNV LNFEGVNGLE QMKWSSPSVD
QVKYDVMIPF IRQVSGPMDY TQGAMRNASK GNYYPCYSEP MSQGTRCRQL ALYVVFESPF
NMLCDTPSNY MREPESTAFI AEIPTVWDES IVLDGKMGEY IVTARRKGDV WYVGGITDWS
ARDIEVDCSF LGDKSYHATL FKDGVNAHRA GRDYKCESFP IKKDGKLKVH LAPGGGFALK
IK
