APU_THEP3
ID APU_THEP3 Reviewed; 1481 AA.
AC P38939; B0K7X5;
DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot.
DT 20-MAY-2008, sequence version 2.
DT 03-AUG-2022, entry version 149.
DE RecName: Full=Amylopullulanase;
DE AltName: Full=Alpha-amylase/pullulanase;
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 {ECO:0000269|PubMed:8344920};
DE AltName: Full=1,4-alpha-D-glucan glucanohydrolase;
DE AltName: Full=Alpha-dextrin endo-1,6-alpha-glucosidase;
DE Flags: Precursor;
GN Name=apu; OrderedLocusNames=Teth39_2173;
OS Thermoanaerobacter pseudethanolicus (strain ATCC 33223 / 39E) (Clostridium
OS thermohydrosulfuricum).
OC Bacteria; Firmicutes; Clostridia; Thermoanaerobacterales;
OC Thermoanaerobacteraceae; Thermoanaerobacter.
OX NCBI_TaxID=340099;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2302196; DOI=10.1016/0006-291x(90)91920-n;
RA Mathupala S.P., Saha B.C., Zeikus J.G.;
RT "Substrate competition and specificity at the active site of
RT amylopullulanase from Clostridium thermohydrosulfuricum.";
RL Biochem. Biophys. Res. Commun. 166:126-132(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 625-630, MUTAGENESIS
RP OF ASP-628; GLU-657 AND ASP-734, CATALYTIC ACTIVITY, AND ACTIVE SITE.
RX PubMed=8344920; DOI=10.1016/s0021-9258(19)85426-1;
RA Mathupala S.P., Lowe S.E., Podkovyrov S.M., Zeikus J.G.;
RT "Sequencing of the amylopullulanase (apu) gene of Thermoanaerobacter
RT ethanolicus 39E, and identification of the active site by site-directed
RT mutagenesis.";
RL J. Biol. Chem. 268:16332-16344(1993).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 33223 / DSM 2355 / 39E;
RA Copeland A., Lucas S., Lapidus A., Barry K., Glavina del Rio T., Dalin E.,
RA Tice H., Pitluck S., Bruce D., Goodwin L., Saunders E., Brettin T.,
RA Detter J.C., Han C., Schmutz J., Larimer F., Land M., Hauser L.,
RA Kyrpides N., Lykidis A., Hemme C., Fields M.W., He Z., Zhou J.,
RA Richardson P.;
RT "Complete sequence of Thermoanaerobacter pseudethanolicus 39E.";
RL Submitted (JAN-2008) to the EMBL/GenBank/DDBJ databases.
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 Evidence={ECO:0000269|PubMed:8344920};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000250|UniProtKB:Q60053};
CC -!- SUBUNIT: Monomer.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=ABY95795.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; M97665; AAA23201.1; -; Genomic_DNA.
DR EMBL; CP000924; ABY95795.1; ALT_FRAME; Genomic_DNA.
DR PIR; S28669; S28669.
DR AlphaFoldDB; P38939; -.
DR SMR; P38939; -.
DR STRING; 340099.Teth39_2173; -.
DR CAZy; CBM20; Carbohydrate-Binding Module Family 20.
DR CAZy; CBM34; Carbohydrate-Binding Module Family 34.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR PRIDE; P38939; -.
DR EnsemblBacteria; ABY95795; ABY95795; Teth39_2173.
DR KEGG; tpd:Teth39_2173; -.
DR eggNOG; COG0366; Bacteria.
DR eggNOG; COG3103; Bacteria.
DR HOGENOM; CLU_002894_0_0_9; -.
DR OMA; WGYDSLP; -.
DR Proteomes; UP000002156; Chromosome.
DR GO; GO:0004556; F:alpha-amylase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
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:UniProtKB-KW.
DR CDD; cd02857; E_set_CDase_PDE_N; 1.
DR CDD; cd00063; FN3; 2.
DR Gene3D; 2.60.40.10; -; 7.
DR Gene3D; 2.60.40.1180; -; 1.
DR InterPro; IPR031319; A-amylase_C.
DR InterPro; IPR013784; Carb-bd-like_fold.
DR InterPro; IPR002044; CBM_fam20.
DR InterPro; IPR003961; FN3_dom.
DR InterPro; IPR036116; FN3_sf.
DR InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR InterPro; IPR004185; Glyco_hydro_13_lg-like_dom.
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; PF02903; Alpha-amylase_N; 1.
DR SMART; SM00642; Aamy; 1.
DR SMART; SM00632; Aamy_C; 1.
DR SMART; SM01065; CBM_2; 1.
DR SMART; SM00060; FN3; 2.
DR SUPFAM; SSF49265; SSF49265; 2.
DR SUPFAM; SSF49452; SSF49452; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF81296; SSF81296; 1.
DR PROSITE; PS51166; CBM20; 1.
DR PROSITE; PS50853; FN3; 2.
PE 1: Evidence at protein level;
KW Calcium; Carbohydrate metabolism; Direct protein sequencing; Glycosidase;
KW Hydrolase; Metal-binding; Reference proteome; Repeat; Signal.
FT SIGNAL 1..31
FT /evidence="ECO:0000255"
FT CHAIN 32..1481
FT /note="Amylopullulanase"
FT /id="PRO_0000001322"
FT DOMAIN 927..1018
FT /note="Fibronectin type-III 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00316"
FT DOMAIN 1159..1255
FT /note="Fibronectin type-III 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00316"
FT DOMAIN 1250..1357
FT /note="CBM20"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00594"
FT REGION 1441..1481
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1452..1474
FT /note="Pro residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 628
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:8344920"
FT ACT_SITE 657
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:8344920"
FT BINDING 245
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 247
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 285
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 340
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 398
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 400
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 404
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 449
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 451
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q60053"
FT BINDING 524
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 MUTAGEN 628
FT /note="D->N,E: Loss of function."
FT /evidence="ECO:0000269|PubMed:8344920"
FT MUTAGEN 657
FT /note="E->Q,D: Loss of function."
FT /evidence="ECO:0000269|PubMed:8344920"
FT MUTAGEN 734
FT /note="D->Q,E: Loss of function."
FT /evidence="ECO:0000269|PubMed:8344920"
FT CONFLICT 185
FT /note="G -> R (in Ref. 1; no nucleotide entry and 2;
FT AAA23201)"
FT /evidence="ECO:0000305"
FT CONFLICT 264..265
FT /note="SP -> FA (in Ref. 1; no nucleotide entry and 2;
FT AAA23201)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 1481 AA; 166230 MW; F9CE0B0BFEF892D3 CRC64;
MFKRRTLGFL LSFLLIYTAV FGSMPVQFAK AETDTAPAIA NVVGDFQSKI GDSDWNINSD
KTVMTYKGNG FYEFTTPVAL PAGDYEYKVA LNHSWEGGGV PSQGNLSLHL DSDSVVTFYY
NYNTSSVTDS TKYTPIPEEK LPRIVGTIQS AIGAGDDWKP ETSTAIMRDY KFNNVYEYTA
NVPKGYYEFK VTLGPSWDIN YGLNGEQNGP NIPLNVAYDT KITFYYDSVS HNIWTDYNPP
LTGPDNNIYY DDLKHDTHDP FFRSPFGAIK TGDTVTLRIQ AKNHDLESAK ISYWDDIKKT
RTEVPMYKIG QSPDGQYEYW EVKLSFDYPT RIWYYFILKD GTKTAYYGDN DEQLGGVGKA
TDTVNKDFEL TVYDKNLDTP DWMKGAVMYQ IFPDRFYNGD PLNDRLKEYS RGFDPVEYHD
DWYDLPDNPN DKDKPGYTGD GIWNNDFFGG DLQGINDKLD YLKNLGISVI YLNPIFQSPS
NHRYDTTDYT KIDELLGDLD TFKTLMKEAH ARGIKVILDG VFNHTSDDSI YFDRYGKYLD
NELGAYQAWK QGDQSKSPYG DWYEIKPDGT YEGWWGFDSL PVIRQINGSE YNVKSWADFI
INNPNAISKY WLNPDGDKDA GADGWRLDVA NEIAHDFWVH FRAAINTVKP NAPMIAELWG
DASLDLLGDS FNSVMNYLFR NAVIDFILDK QFDDGNVVHN PIDAAKLDQR LMSIYERYPL
PVFYSTMNLL GSHDTMRILT VFGYNSANEN QNSQEAKDLA VKRLKLAAIL QMGYPGMPSI
YYGDEAGQSG GKDPDNRRTF SWGREDKDLQ DFFKKVVNIR NENQVLKTGD LETLYANGDV
YAFGRRIING KDVFGNSYPD SVAIVVINKG EAKSVQIDTT KFVRDGVAFT DALSGKTYTV
RDGQIVVEVV ALDGAILISD PGQNLTAPQP ITDLKAVSGN GQVDLSWSAV DRAVSYNIYR
STVKGGLYEK IASNVTQITY IDTDVTNGLK YVYSVTAVDS DGNESALSNE VEAYPAFSIG
WAGNMNQVDT HVIGVNNPVE VYAEIWAEGL TDKPGQGENM IAQLGYRYIG DGGQDATRNK
VEGVEINKDW TWVDARYVGD SGNNDKYMAK FVPDMVGTWE YIMRFSSNQG QDWTYTKGPD
GKTDEAKQFI VVPSNDVEPP TALGLQQPGI ESSRVTLNWS LSTDNVAIYG YEIYKSLSET
GPFVKIATVA DTVYNYVDTD VVNGKVYYYK VVAVDTSFNR TASNIVKATP DIIPIKVIFN
VTVPDYTPDD GANIAGNFHD AFWNPSAHQM TKTGPNTYSI TLTLNEGTQL EYKYARGSWD
KVEKGEYGEE IANRKITVVN QGSNTMVVND TVQRWRDLPI YIYSPKDNTT VDANTNEIEI
KGNTYKGAKV TINDESFVQQ ENGVFTKVVP LEYGVNTTKI HVEPSGDKNN ELTKDITITV
IREEPVQEKE PTPTPESEPA PMPEPQPTPT PEPQPSAIMA L
