ID   APU_THETU               Reviewed;        1861 AA.
AC   P38536;
DT   01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1995, sequence version 2.
DT   02-SEP-2026, entry version 143.
DE   RecName: Full=Amylopullulanase;
DE   AltName: Full=Alpha-amylase/pullulanase;
DE   AltName: Full=Pullulanase type II;
DE   Includes:
DE     RecName: Full=Alpha-amylase;
DE              EC=3.2.1.1;
DE     AltName: Full=1,4-alpha-D-glucan glucanohydrolase;
DE   Includes:
DE     RecName: Full=Pullulanase;
DE              EC=3.2.1.41;
DE     AltName: Full=1,4-alpha-D-glucan glucanohydrolase;
DE     AltName: Full=Alpha-dextrin endo-1,6-alpha-glucosidase;
DE   Flags: Precursor;
GN   Name=amyB;
OS   Thermoanaerobacterium thermosulfurigenes (Clostridium thermosulfurogenes).
OC   Bacteria; Bacillati; Bacillota; Clostridia; Thermoanaerobacterales;
OC   Thermoanaerobacteraceae; Thermoanaerobacterium.
OX   NCBI_TaxID=33950;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=DSM 3896 / EM1;
RX   PubMed=8195085; DOI=10.1128/jb.176.11.3295-3302.1994;
RA   Matuschek M., Burchhardt G., Sahm K., Bahl H.;
RT   "Pullulanase of Thermoanaerobacterium thermosulfurigenes EM1 (Clostridium
RT   thermosulfurogenes): molecular analysis of the gene, composite structure of
RT   the enzyme, and a common model for its attachment to the cell surface.";
RL   J. Bacteriol. 176:3295-3302(1994).
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Endohydrolysis of (1->4)-alpha-D-glucosidic linkages in
CC         polysaccharides containing three or more (1->4)-alpha-linked D-
CC         glucose units.; EC=3.2.1.1;
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Hydrolysis of (1->6)-alpha-D-glucosidic linkages in pullulan,
CC         amylopectin and glycogen, and in the alpha- and beta-limit dextrins
CC         of amylopectin and glycogen.; EC=3.2.1.41;
CC   -!- COFACTOR:
CC       Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC         Evidence={ECO:0000250|UniProtKB:Q60053};
CC   -!- SUBCELLULAR LOCATION: Secreted, cell wall. Note=It C-terminus may serve
CC       as an S-layer anchor.
CC   -!- PTM: Glycosylated.
CC   -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR   EMBL; M57692; AAB00841.1; -; Genomic_DNA.
DR   AlphaFoldDB; P38536; -.
DR   SMR; P38536; -.
DR   CAZy; CBM20; Carbohydrate-Binding Module Family 20.
DR   CAZy; CBM34; Carbohydrate-Binding Module Family 34.
DR   CAZy; GH13; Glycoside Hydrolase Family 13.
DR   GO; GO:0004556; F:alpha-amylase activity; IEA:UniProtKB-EC.
DR   GO; GO:0051060; F:pullulanase activity; IEA:UniProtKB-EC.
DR   GO; GO:2001070; F:starch binding; IEA:InterPro.
DR   GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR   CDD; cd11338; AmyAc_CMD; 1.
DR   CDD; cd02857; E_set_CDase_PDE_N; 1.
DR   CDD; cd00063; FN3; 1.
DR   CDD; cd12962; X25_BaPul_like; 2.
DR   Gene3D; 3.20.20.80; Glycosidases; 1.
DR   Gene3D; 2.60.40.1180; Golgi alpha-mannosidase II; 1.
DR   Gene3D; 2.60.40.10; Immunoglobulins; 6.
DR   InterPro; IPR031319; A-amylase_C.
DR   InterPro; IPR013784; Carb-bd-like_fold.
DR   InterPro; IPR002044; CBM20.
DR   InterPro; IPR003961; FN3_dom.
DR   InterPro; IPR036116; FN3_sf.
DR   InterPro; IPR006047; GH13_cat_dom.
DR   InterPro; IPR017853; GH_hydrolase_sf.
DR   InterPro; IPR004185; Glyco_hydro_13_lg-like_dom.
DR   InterPro; IPR013780; Glyco_hydro_b.
DR   InterPro; IPR013783; Ig-like_fold.
DR   InterPro; IPR014756; Ig_E-set.
DR   InterPro; IPR001119; SLH_dom.
DR   InterPro; IPR054409; X25_BaPul-like.
DR   PANTHER; PTHR10357; ALPHA-AMYLASE FAMILY MEMBER; 1.
DR   PANTHER; PTHR10357:SF210; MALTODEXTRIN GLUCOSIDASE; 1.
DR   Pfam; PF00128; Alpha-amylase; 1.
DR   Pfam; PF02903; Alpha-amylase_N; 1.
DR   Pfam; PF00395; SLH; 3.
DR   Pfam; PF22058; X25_BaPul_like; 2.
DR   SMART; SM00642; Aamy; 1.
DR   SMART; SM00632; Aamy_C; 1.
DR   SMART; SM01065; CBM_2; 1.
DR   SMART; SM00060; FN3; 2.
DR   SUPFAM; SSF51445; (Trans)glycosidases; 1.
DR   SUPFAM; SSF81296; E set domains; 1.
DR   SUPFAM; SSF49265; Fibronectin type III; 2.
DR   SUPFAM; SSF51011; Glycosyl hydrolase domain; 1.
DR   SUPFAM; SSF49452; Starch-binding domain-like; 1.
DR   PROSITE; PS51166; CBM20; 1.
DR   PROSITE; PS50853; FN3; 2.
DR   PROSITE; PS51272; SLH; 3.
PE   3: Inferred from homology;
KW   Calcium; Carbohydrate metabolism; Cell wall; Glycoprotein; Glycosidase;
KW   Hydrolase; Metal-binding; Repeat; Secreted; Signal.
FT   SIGNAL          1..35
FT                   /evidence="ECO:0000255"
FT   CHAIN           36..1861
FT                   /note="Amylopullulanase"
FT                   /id="PRO_0000001325"
FT   DOMAIN          929..1021
FT                   /note="Fibronectin type-III 1"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00316"
FT   DOMAIN          1158..1252
FT                   /note="Fibronectin type-III 2"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00316"
FT   DOMAIN          1246..1354
FT                   /note="CBM20"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00594"
FT   DOMAIN          1677..1740
FT                   /note="SLH 1"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00777"
FT   DOMAIN          1741..1799
FT                   /note="SLH 2"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00777"
FT   DOMAIN          1802..1861
FT                   /note="SLH 3"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00777"
FT   REGION          1448..1486
FT                   /note="Disordered"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   COMPBIAS        1450..1486
FT                   /note="Low complexity"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   ACT_SITE        628
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   ACT_SITE        657
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         248
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         250
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         288
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         343
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         401
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         403
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         406
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         407
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         453
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000250|UniProtKB:Q60053"
FT   BINDING         526
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:P38940"
FT   BINDING         626
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:P38940"
FT   BINDING         733..734
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:P38940"
FT   BINDING         793
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:P38940"
FT   BINDING         797
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:P38940"
FT   SITE            734
FT                   /note="Transition state stabilizer"
FT                   /evidence="ECO:0000250"
FT   CONFLICT        1734
FT                   /note="D -> E (in Ref. 1; AAB00841)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   1861 AA;  206104 MW;  06C23070E453B574 CRC64;
     MNKKLFTNRF ISFNMSLLLV LTAVFSSIPL HSVHAADNAS VVANIVGEFQ DQLGDSNWNI
     DSNITIMQYM GNGLYEFTTP TQLKAGSYQY KVALNHSWDG GGVPSQGNLT LNLANDSYVT
     FWFDYNTQSV TDSTKYTPIA NDKLPRLVGT IQSAIGAGND WKPETSTAIM TDDNFDNVYS
     YTAHVPKGDY QYKVTLGNTW DENYGANGVK DGSNIQINVT NDADITFYYD ANTHNIWTNY
     SPILTGLDNN IYYDDLKHDT HDSFFRNPFG AVKVDQTVTL RIQAKNHDLE SARISYWDDI
     NKIRIELPMT RIGESPDGNF EYWEIKLSFD HPTRIWYYFI LKDGTKTAYY GDNDDQLGGV
     GKATDTVNKD FELTVYDKNF DTPDWMKGAV MYQIFPDRFY NGDTSNDHAK TLSRGNDPIE
     FHNNWNDLPD NPNNAGTPGY TGDGIWSNDF FGDLKGIDDK LDYLKGLGVS VIYLNPIFES
     PSNHKYDTAD YTKIDEMFGT TQDFEKLMSD AHAKGIKIIL DGVFNHTSDD SIYFNRYGKY
     PGLGAYQAWK EGNQSLSPYG DWYTINSDGT YECWWGYDSL PVIKSLNGSE YNVTSWANFI
     INDENAISKY WLNPDGNLND GADGWRLDVE NEVAHDFWTH FRNAINTVKF EAPMIAENWG
     DASLDLLGDS FNSVMNYQFR NDIIDFLIGQ SFDDGNGQHN PIDAAKLDQR LMSIYERYPL
     PAFYSTMNLL GSHDTMRILT VFGYNSADPN ENSDAAKQLA EQKLKLATIL QMGYPGMADI
     YYGDEAGVSG GKDPDDRRTF PWGNEDTTLQ DFFKNISSIR NNNQVLKTGD LETLYAQNDV
     YAIGRRIING KDAFGTSYPD SAAIVAINRS KSDKQIAIDT TKFLRDGVTF KDLINNNVSY
     SISNGQIVID VPAMSGVMLI SDDGQDLTAP QAPSNVVVTS GNGKVDLSWL QSDGATGYNI
     YRSSVEGGLY EKIASNVTET TFEDANVTNG LKYVYAISAI DELGNESGIS NDAVAYPAYP
     IGWVGNLTQV SDNHIIGVDK PTEDIYAEVW ADGLTNSTGQ GPNMIAQLGY KYVSGTVYDS
     VYGSVYNSVY GVDDSGFTWV NAQYVGDIGN NDQYKASFTP DKIGQWEYLM RFSDNQGQDW
     ITTSTLSFYV VPSDDLIKPT APYLNQPGTE SSRVSLTWNP STDNVGIYDY EIYRSDGGTF
     NKIATVSNEV YNYIDTSVIN GVTYNYKVVA VDLSFNRTES NVVTIKPDVV PIKVIFNVTV
     PDYTPDAVNL AGTFPNATWD PSAQQMTKID NNTYSITLTL DEGTQIEYKY ARGSWDKVEK
     DEYGNEFASN RKVTIVNQGN NEMTINDTVY RWRDIPIFIY SPSSNMTVDS NISTMEVKGN
     TYKGAKVTIN GDSFVQDKNG VFTKDVSLNY GVNKIKIHVE PNDGSVYGND QGRITELTKD
     IEIDVIRQEN NSGSGTGNNN TSTSGSNSSS TGSGSTGSTS ITSNISNTSN TSNTIGVITK
     NGNVITLTLD AGKAKDLIVN SKDKKVVFDI TTIGEGQQKV VQISKDILDT SAANGKDIVI
     KSDNASIALT KDALNQNQIQ NGVNVSIKDN GKPNVTNYVS LSNVVDITIS GISGNVTLAK
     PVEVTLNISK ANDPRKVAVY YYNPTTNQWE YVGGKVDASS GTITFNATHF SQYAAFEYDK
     TFNDIKDNWA KDVIEVLASR HIVEGMTDTQ YEPNKTVTRA EFTAMILRLL NIKDETYSGE
     FSDVKSGDWY ANAIEAAYKA GIIEGDGKNA RPNDSITREE MTAIAMRAYE MLTQYKEENI
     GATTFSDDKS ISDWARNVVA NAAKLGIVNG EPNNVFAPKG NATRAEAAAI IYGLLEKSGN
     I
//