AMYG_ASPAW
ID AMYG_ASPAW Reviewed; 640 AA.
AC P69327; P04064; Q92201; Q99179;
DT 01-NOV-1986, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1986, sequence version 1.
DT 03-AUG-2022, entry version 90.
DE RecName: Full=Glucoamylase;
DE EC=3.2.1.3;
DE AltName: Full=1,4-alpha-D-glucan glucohydrolase;
DE AltName: Full=Glucan 1,4-alpha-glucosidase;
DE Flags: Precursor;
GN Name=GLAA;
OS Aspergillus awamori (Black koji mold).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus.
OX NCBI_TaxID=105351;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS G1 AND G2).
RX PubMed=6440004; DOI=10.1128/mcb.4.11.2306-2315.1984;
RA Nunberg J.H., Meade J.H., Cole G., Lawyer F.C., McCabe P., Schweickart V.,
RA Tal R., Wittman V.P., Flatgaard J.E., Innis M.A.;
RT "Molecular cloning and characterization of the glucoamylase gene of
RT Aspergillus awamori.";
RL Mol. Cell. Biol. 4:2306-2315(1984).
RN [2]
RP SEQUENCE REVISION.
RA Nunberg J.H., Meade J.H., Cole G., Lawyer F.C., McCabe P., Schweickart V.,
RA Tal R., Wittman V.P., Flatgaard J.E., Innis M.A.;
RL Submitted (FEB-1985) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP ACTIVE SITES, AND MUTAGENESIS.
RX PubMed=1970434; DOI=10.1093/protein/3.3.193;
RA Sierks M.R., Ford C., Reilly P.J., Svensson B.;
RT "Catalytic mechanism of fungal glucoamylase as defined by mutagenesis of
RT Asp176, Glu179 and Glu180 in the enzyme from Aspergillus awamori.";
RL Protein Eng. 3:193-198(1990).
RN [4]
RP MUTAGENESIS OF TRP-144.
RX PubMed=2510150; DOI=10.1093/protein/2.8.621;
RA Sierks M.R., Ford C., Reilly P.J., Svensson B.;
RT "Site-directed mutagenesis at the active site Trp120 of Aspergillus awamori
RT glucoamylase.";
RL Protein Eng. 2:621-625(1989).
RN [5]
RP MUTAGENESIS.
RX PubMed=8433972; DOI=10.1093/protein/6.1.75;
RA Sierks M.R., Ford C., Reilly P.J., Svensson B.;
RT "Functional roles and subsite locations of Leu177, Trp178 and Asn182 of
RT Aspergillus awamori glucoamylase determined by site-directed mutagenesis.";
RL Protein Eng. 6:75-79(1993).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, GLYCOSYLATION, AND
RP DISULFIDE BONDS.
RC STRAIN=Var. X100;
RX PubMed=1527049; DOI=10.2210/pdb1gly/pdb;
RA Aleshin A., Golubev A., Firsov L.M., Honzatko R.B.;
RT "Crystal structure of glucoamylase from Aspergillus awamori var. X100 to
RT 2.2-A resolution.";
RL J. Biol. Chem. 267:19291-19298(1992).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 25-495.
RC STRAIN=Var. X100;
RX PubMed=8195212; DOI=10.1016/s0021-9258(17)40728-9;
RA Aleshin A., Firsov L.M., Honzatko R.B.;
RT "Refined structure for the complex of acarbose with glucoamylase from
RT Aspergillus awamori var. X100 to 2.4-A resolution.";
RL J. Biol. Chem. 269:15631-15639(1994).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, AND GLYCOSYLATION.
RC STRAIN=Var. X100;
RX PubMed=8176747; DOI=10.1006/jmbi.1994.1316;
RA Aleshin A., Hoffman C., Firsov L.M., Honzatko R.B.;
RT "Refined crystal structures of glucoamylase from Aspergillus awamori var.
RT X100.";
RL J. Mol. Biol. 238:575-591(1994).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal (1->4)-linked alpha-D-glucose residues
CC successively from non-reducing ends of the chains with release of
CC beta-D-glucose.; EC=3.2.1.3;
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative splicing; Named isoforms=2;
CC Name=G1;
CC IsoId=P69327-1; Sequence=Displayed;
CC Name=G2;
CC IsoId=P69327-2; Sequence=VSP_012836, VSP_000262;
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 15 family. {ECO:0000305}.
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DR EMBL; K02465; AAB59296.1; -; mRNA.
DR EMBL; K02465; AAB59297.1; -; mRNA.
DR PIR; A29166; A29166.
DR PIR; A93066; A29776.
DR PDB; 1AGM; X-ray; 2.30 A; A=25-495.
DR PDB; 1DOG; X-ray; 2.30 A; A=25-495.
DR PDB; 1GAH; X-ray; 2.00 A; A=25-496.
DR PDB; 1GAI; X-ray; 1.70 A; A=25-497.
DR PDB; 1GLM; X-ray; 2.40 A; A=25-495.
DR PDB; 3GLY; X-ray; 2.20 A; A=25-495.
DR PDBsum; 1AGM; -.
DR PDBsum; 1DOG; -.
DR PDBsum; 1GAH; -.
DR PDBsum; 1GAI; -.
DR PDBsum; 1GLM; -.
DR PDBsum; 3GLY; -.
DR AlphaFoldDB; P69327; -.
DR BMRB; P69327; -.
DR SMR; P69327; -.
DR CAZy; CBM20; Carbohydrate-Binding Module Family 20.
DR CAZy; GH15; Glycoside Hydrolase Family 15.
DR GlyConnect; 174; 2 N-Linked glycans (2 sites).
DR BRENDA; 3.2.1.3; 494.
DR EvolutionaryTrace; P69327; -.
DR GO; GO:0004339; F:glucan 1,4-alpha-glucosidase activity; IEA:UniProtKB-EC.
DR GO; GO:2001070; F:starch binding; IEA:InterPro.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR CDD; cd05811; CBM20_glucoamylase; 1.
DR Gene3D; 1.50.10.10; -; 1.
DR Gene3D; 2.60.40.10; -; 1.
DR InterPro; IPR008928; 6-hairpin_glycosidase_sf.
DR InterPro; IPR012341; 6hp_glycosidase-like_sf.
DR InterPro; IPR013784; Carb-bd-like_fold.
DR InterPro; IPR034836; CBM20_glucoamylase.
DR InterPro; IPR002044; CBM_fam20.
DR InterPro; IPR011613; GH15-like.
DR InterPro; IPR000165; Glucoamylase.
DR InterPro; IPR008291; Glucoamylase_SBD.
DR InterPro; IPR013783; Ig-like_fold.
DR Pfam; PF00686; CBM_20; 1.
DR Pfam; PF00723; Glyco_hydro_15; 1.
DR PIRSF; PIRSF001031; Glu-a-glcsd_SBD; 1.
DR PRINTS; PR00736; GLHYDRLASE15.
DR SMART; SM01065; CBM_2; 1.
DR SUPFAM; SSF48208; SSF48208; 1.
DR SUPFAM; SSF49452; SSF49452; 1.
DR PROSITE; PS51166; CBM20; 1.
DR PROSITE; PS00820; GLUCOAMYLASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alternative splicing; Carbohydrate metabolism;
KW Cleavage on pair of basic residues; Disulfide bond; Glycoprotein;
KW Glycosidase; Hydrolase; Polysaccharide degradation; Signal.
FT SIGNAL 1..18
FT /evidence="ECO:0000255"
FT PROPEP 19..24
FT /id="PRO_0000001459"
FT CHAIN 25..640
FT /note="Glucoamylase"
FT /id="PRO_0000001460"
FT DOMAIN 533..640
FT /note="CBM20"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00594"
FT REGION 498..533
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 616..640
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 623..640
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 200
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10051,
FT ECO:0000269|PubMed:1970434"
FT ACT_SITE 203
FT /note="Proton donor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10051,
FT ECO:0000269|PubMed:1970434"
FT BINDING 144
FT /ligand="substrate"
FT CARBOHYD 195
FT /note="N-linked (GlcNAc...) asparagine"
FT /id="CAR_000112"
FT CARBOHYD 419
FT /note="N-linked (GlcNAc...) asparagine"
FT /id="CAR_000113"
FT CARBOHYD 465
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 467
FT /note="O-linked (Man) serine"
FT CARBOHYD 468
FT /note="O-linked (Man) serine"
FT CARBOHYD 476
FT /note="O-linked (Man) threonine"
FT CARBOHYD 477
FT /note="O-linked (Man) serine"
FT CARBOHYD 483
FT /note="O-linked (Man) serine"
FT CARBOHYD 484
FT /note="O-linked (Man) serine"
FT CARBOHYD 486
FT /note="O-linked (Man) threonine"
FT CARBOHYD 488
FT /note="O-linked (Man) threonine"
FT CARBOHYD 492
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 496
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 499
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 500
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 501
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 502
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 504
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 506
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 508
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 510
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 512
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 513
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 514
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 515
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 517
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 518
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 520
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 522
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 524
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 525
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 526
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 527
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 528
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 529
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 530
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 531
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 532
FT /note="O-linked (Man) serine"
FT /evidence="ECO:0000250"
FT CARBOHYD 534
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT CARBOHYD 535
FT /note="O-linked (Man) threonine"
FT /evidence="ECO:0000250"
FT DISULFID 234..237
FT /evidence="ECO:0000269|PubMed:1527049"
FT DISULFID 246..473
FT /evidence="ECO:0000269|PubMed:1527049"
FT DISULFID 286..294
FT /evidence="ECO:0000269|PubMed:1527049"
FT VAR_SEQ 527..534
FT /note="STSSTSCT -> TTRSGMSL (in isoform G2)"
FT /evidence="ECO:0000303|PubMed:6440004"
FT /id="VSP_012836"
FT VAR_SEQ 535..640
FT /note="Missing (in isoform G2)"
FT /evidence="ECO:0000303|PubMed:6440004"
FT /id="VSP_000262"
FT MUTAGEN 144
FT /note="W->Y: 2% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:2510150"
FT HELIX 26..44
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 47..49
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 67..71
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 74..76
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 77..92
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 96..98
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 99..114
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 122..125
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 127..129
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 150..168
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 172..177
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 179..193
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 206..209
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 210..229
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 235..248
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 249..251
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 254..257
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 259..262
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 271..277
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 288..291
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 296..307
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 308..312
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 314..316
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 335..337
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 342..362
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 364..367
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 369..371
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 372..378
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 384..388
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 392..415
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 424..426
FT /evidence="ECO:0007829|PDB:1GAI"
FT TURN 428..430
FT /evidence="ECO:0007829|PDB:1GAI"
FT STRAND 433..438
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 440..453
FT /evidence="ECO:0007829|PDB:1GAI"
FT HELIX 463..465
FT /evidence="ECO:0007829|PDB:1GAI"
SQ SEQUENCE 640 AA; 68309 MW; 26F58DD18AD7F702 CRC64;
MSFRSLLALS GLVCTGLANV ISKRATLDSW LSNEATVART AILNNIGADG AWVSGADSGI
VVASPSTDNP DYFYTWTRDS GLVLKTLVDL FRNGDTSLLS TIENYISAQA IVQGISNPSG
DLSSGAGLGE PKFNVDETAY TGSWGRPQRD GPALRATAMI GFGQWLLDNG YTSTATDIVW
PLVRNDLSYV AQYWNQTGYD LWEEVNGSSF FTIAVQHRAL VEGSAFATAV GSSCSWCDSQ
APEILCYLQS FWTGSFILAN FDSSRSGKDA NTLLGSIHTF DPEAACDDST FQPCSPRALA
NHKEVVDSFR SIYTLNDGLS DSEAVAVGRY PEDTYYNGNP WFLCTLAAAE QLYDALYQWD
KQGSLEVTDV SLDFFKALYS DAATGTYSSS SSTYSSIVDA VKTFADGFVS IVETHAASNG
SMSEQYDKSD GEQLSARDLT WSYAALLTAN NRRNSVVPAS WGETSASSVP GTCAATSAIG
TYSSVTVTSW PSIVATGGTT TTATPTGSGS VTSTSKTTAT ASKTSTSTSS TSCTTPTAVA
VTFDLTATTT YGENIYLVGS ISQLGDWETS DGIALSADKY TSSDPLWYVT VTLPAGESFE
YKFIRIESDD SVEWESDPNR EYTVPQACGT STATVTDTWR
