ID   ISOA_FLASP              Reviewed;         777 AA.
AC   O32611;
DT   15-DEC-1998, integrated into UniProtKB/Swiss-Prot.
DT   01-JAN-1998, sequence version 1.
DT   03-AUG-2022, entry version 92.
DE   RecName: Full=Isoamylase;
DE            EC=3.2.1.68;
DE   Flags: Precursor;
GN   Name=iam;
OS   Flavobacterium sp.
OC   Bacteria; Bacteroidetes; Flavobacteriia; Flavobacteriales;
OC   Flavobacteriaceae; Flavobacterium; unclassified Flavobacterium.
OX   NCBI_TaxID=239;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX   PubMed=9197405; DOI=10.1007/s004380050441;
RA   Krohn B.M., Barry G.F., Kishore G.M.;
RT   "An isoamylase with neutral pH optimum from a Flavobacterium species:
RT   cloning, characterization and expression of the iam gene.";
RL   Mol. Gen. Genet. 254:469-478(1997).
CC   -!- FUNCTION: Has a high rate of hydrolysis for glycogen. Does not cleave
CC       pullulan.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Hydrolysis of (1->6)-alpha-D-glucosidic branch linkages in
CC         glycogen, amylopectin and their beta-limit dextrins.; EC=3.2.1.68;
CC   -!- COFACTOR:
CC       Name=Ca(2+); Xref=ChEBI:CHEBI:29108; Evidence={ECO:0000250};
CC       Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000250};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       pH dependence:
CC         Optimum pH is 6-7.;
CC   -!- SUBUNIT: Monomer. {ECO:0000250}.
CC   -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR   EMBL; U90120; AAB63356.1; -; Genomic_DNA.
DR   AlphaFoldDB; O32611; -.
DR   SMR; O32611; -.
DR   CAZy; CBM48; Carbohydrate-Binding Module Family 48.
DR   CAZy; GH13; Glycoside Hydrolase Family 13.
DR   GO; GO:0004133; F:glycogen debranching enzyme activity; IEA:UniProt.
DR   GO; GO:0019156; F:isoamylase activity; IEA:UniProtKB-EC.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   CDD; cd02856; E_set_GDE_Isoamylase_N; 1.
DR   Gene3D; 2.60.40.10; -; 1.
DR   Gene3D; 2.60.40.1180; -; 1.
DR   InterPro; IPR044505; GlgX_Isoamylase_N_E_set.
DR   InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR   InterPro; IPR004193; Glyco_hydro_13_N.
DR   InterPro; IPR013780; Glyco_hydro_b.
DR   InterPro; IPR017853; Glycoside_hydrolase_SF.
DR   InterPro; IPR013783; Ig-like_fold.
DR   InterPro; IPR014756; Ig_E-set.
DR   Pfam; PF00128; Alpha-amylase; 1.
DR   Pfam; PF02922; CBM_48; 1.
DR   SMART; SM00642; Aamy; 1.
DR   SUPFAM; SSF51445; SSF51445; 1.
DR   SUPFAM; SSF81296; SSF81296; 1.
PE   1: Evidence at protein level;
KW   Calcium; Disulfide bond; Glycosidase; Hydrolase; Metal-binding; Signal.
FT   SIGNAL          1..32
FT                   /evidence="ECO:0000255"
FT   CHAIN           33..777
FT                   /note="Isoamylase"
FT                   /id="PRO_0000001423"
FT   ACT_SITE        410
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000250"
FT   ACT_SITE        458
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000250"
FT   BINDING         162
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /evidence="ECO:0000250"
FT   BINDING         263
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /evidence="ECO:0000250"
FT   BINDING         264
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /evidence="ECO:0000250"
FT   BINDING         266
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /evidence="ECO:0000250"
FT   BINDING         293
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /evidence="ECO:0000250"
FT   SITE            533
FT                   /note="Transition state stabilizer"
FT                   /evidence="ECO:0000250"
FT   DISULFID        419..423
FT                   /evidence="ECO:0000250"
SQ   SEQUENCE   777 AA;  84340 MW;  90548988EFD1445B CRC64;
     MDPHAPQRQR SGQRLRALAL AALACALSPA HAAIDAQQLG ARYDAAQANL AFRVYSSRAT
     RVEVFLYKNP TGSQEVARLA LSKDPATQVW SLSLPTSTIK NTYGITGAVY YGYRAWGPNW
     PYDAAWTKGS ATGFVSDVDN AGNRFNPNKL LLDPYAREIS QDPNTATCAD GTIYATGAAH
     RNKDSGLCAS KGIALAADAT SVGSKPTRAL KDEVIYEVHV RGLTRNDDSV PAAERGTYKG
     AARKAAALAA LGVTAVEFLP VQETQNDQND VDPNSTAGDN YWGYMTLNYF APDRRYAYDK
     SAGGPTREWK AMVKAFHDAG IKVYIDVVYN HTGEGGPWSG TDGLSVYNLL SFRGLDNPAY
     YSLSSDYKYP WDNTGVGGNY NTRHPIAQNL IVDSLAYWRD ALGVDGFRFD LASVLGNSCQ
     HGCFNFDKND SGNALNRIVA ELPPRPAAGG AGADLIAEPW AIGGNSYQVG GFPAGWAEWN
     GLYRDALRKK QNKLGVETVT PGTLATRFAG SNDLYGDDGR KPWHSINFVV AHDGFTLNDL
     YAYNDKQNNQ PWPYGPSDGG EDHNLSWNQG GIVAEQRKAA RTGLALLMLS AGVPMITGGD
     EALRTQFGNN NTYNLDSAAN WLYWSRSALE ADHETYTKRL IAFRKAHPAL RPANFYSASD
     TNGNVMEQLR WFKPDGAQAD SAYFNGADNH ALAWRIDGSE FGDSASAIYV AYNGWSGAVD
     FKLPWPGTGK QWYRVTDTAT WNEGPNAVAL PGSETLIGGE NTVYGMQARS LLLLIAK