NEPU1_THEVU
ID NEPU1_THEVU Reviewed; 666 AA.
AC Q60053;
DT 15-AUG-2003, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 108.
DE RecName: Full=Neopullulanase 1;
DE EC=3.2.1.135 {ECO:0000269|PubMed:7548164};
DE AltName: Full=Alpha-amylase I;
DE AltName: Full=TVA I;
DE Flags: Precursor;
GN Name=tvaI;
OS Thermoactinomyces vulgaris.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Thermoactinomycetaceae;
OC Thermoactinomyces.
OX NCBI_TaxID=2026;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CHARACTERIZATION, CATALYTIC ACTIVITY,
RP AND FUNCTION.
RC STRAIN=R-47;
RX PubMed=7548164; DOI=10.1016/0167-4838(95)00101-y;
RA Tonozuka T., Mogi S., Shimura Y., Ibuka A., Sakai H., Matsuzawa H.,
RA Sakano Y., Ohta T.;
RT "Comparison of primary structures and substrate specificities of two
RT pullulan-hydrolyzing alpha-amylases, TVA I and TVA II, from
RT Thermoactinomyces vulgaris R-47.";
RL Biochim. Biophys. Acta 1252:35-42(1995).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF 30-666 IN COMPLEX WITH CALCIUM,
RP AND COFACTOR.
RC STRAIN=R-47;
RX PubMed=12051850; DOI=10.1016/s0022-2836(02)00111-0;
RA Kamitori S., Abe A., Ohtaki A., Kaji A., Tonozuka T., Sakano Y.;
RT "Crystal structures and structural comparison of Thermoactinomyces vulgaris
RT R-47 alpha-amylase 1 (TVAI) at 1.6 A resolution and alpha-amylase 2 (TVAII)
RT at 2.3 A resolution.";
RL J. Mol. Biol. 318:443-453(2002).
CC -!- FUNCTION: Endohydrolysis of 1,4-alpha-glucosidic linkages in pullulan
CC to form panose. Also hydrolyzes cyclodextrins.
CC {ECO:0000269|PubMed:7548164}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of pullulan to panose (6-alpha-D-glucosylmaltose).;
CC EC=3.2.1.135; Evidence={ECO:0000269|PubMed:7548164};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:12051850};
CC Note=Binds 3 Ca(2+) ions per subunit. {ECO:0000269|PubMed:12051850};
CC -!- SUBCELLULAR LOCATION: Secreted.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR EMBL; D13177; BAA02471.1; -; Genomic_DNA.
DR PDB; 1IZJ; X-ray; 2.20 A; A=30-666.
DR PDB; 1IZK; X-ray; 2.20 A; A=30-666.
DR PDB; 1JI1; X-ray; 1.60 A; A/B=30-666.
DR PDB; 1UH2; X-ray; 2.00 A; A=30-666.
DR PDB; 1UH3; X-ray; 2.60 A; A=30-666.
DR PDB; 1UH4; X-ray; 1.80 A; A=30-666.
DR PDB; 2D0F; X-ray; 2.08 A; A=30-666.
DR PDB; 2D0G; X-ray; 2.60 A; A=30-666.
DR PDB; 2D0H; X-ray; 2.10 A; A=30-666.
DR PDB; 5Z0T; X-ray; 1.50 A; A/B=30-666.
DR PDB; 5Z0U; X-ray; 1.37 A; A=30-666.
DR PDBsum; 1IZJ; -.
DR PDBsum; 1IZK; -.
DR PDBsum; 1JI1; -.
DR PDBsum; 1UH2; -.
DR PDBsum; 1UH3; -.
DR PDBsum; 1UH4; -.
DR PDBsum; 2D0F; -.
DR PDBsum; 2D0G; -.
DR PDBsum; 2D0H; -.
DR PDBsum; 5Z0T; -.
DR PDBsum; 5Z0U; -.
DR AlphaFoldDB; Q60053; -.
DR SMR; Q60053; -.
DR DrugBank; DB01841; 4,6-Dideoxyglucose.
DR DrugBank; DB02120; 6-Amino-4-Hydroxymethyl-Cyclohex-4-Ene-1,2,3-Triol.
DR DrugBank; DB02379; Beta-D-Glucose.
DR CAZy; CBM34; Carbohydrate-Binding Module Family 34.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR EvolutionaryTrace; Q60053; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0031216; F:neopullulanase activity; IEA:UniProtKB-EC.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW.
DR CDD; cd02857; E_set_CDase_PDE_N; 1.
DR Gene3D; 2.60.40.10; -; 1.
DR Gene3D; 2.60.40.1180; -; 1.
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 SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF81296; SSF81296; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Chloride; Glycosidase;
KW Hydrolase; Metal-binding; Secreted; Signal.
FT SIGNAL 1..29
FT CHAIN 30..666
FT /note="Neopullulanase 1"
FT /id="PRO_0000001445"
FT ACT_SITE 385
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT ACT_SITE 425
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 31
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 33
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 35
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 71
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 125
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 174
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 176
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 179
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 180
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 216
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 218
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 296
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 305
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 309
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 310
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 312
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 317
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI1"
FT BINDING 383
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 500..501
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 545
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 549
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT SITE 501
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250"
FT STRAND 32..34
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 38..40
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 47..49
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 61..68
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 73..81
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 82..85
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 86..91
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 93..97
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 101..110
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 117..125
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 128..133
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 136..140
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 147..149
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 157..161
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 164..167
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 169..172
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 177..179
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 187..190
FT /evidence="ECO:0007829|PDB:1IZK"
FT HELIX 210..212
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 219..224
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 226..230
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 231..233
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 237..241
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 247..250
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 255..260
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 262..264
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 267..278
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 280..284
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 287..292
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 302..304
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 310..312
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 315..317
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 324..326
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 329..331
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 332..334
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 339..341
FT /evidence="ECO:0007829|PDB:5Z0T"
FT STRAND 345..347
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 356..361
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 368..373
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 375..377
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 381..384
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 387..389
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 397..399
FT /evidence="ECO:0007829|PDB:1UH2"
FT HELIX 403..416
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 421..424
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 431..433
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 434..436
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 437..439
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 441..444
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 446..449
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 450..458
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 471..482
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 487..492
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 494..497
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 505..508
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 509..511
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 513..525
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 526..533
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 536..538
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 545..548
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 554..556
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 562..576
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 578..582
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 583..591
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 592..595
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 596..602
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 607..613
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 615..617
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 619..623
FT /evidence="ECO:0007829|PDB:5Z0U"
FT HELIX 625..628
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 635..638
FT /evidence="ECO:0007829|PDB:5Z0U"
FT TURN 639..641
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 644..646
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 651..656
FT /evidence="ECO:0007829|PDB:5Z0U"
FT STRAND 660..666
FT /evidence="ECO:0007829|PDB:5Z0U"
SQ SEQUENCE 666 AA; 74296 MW; E99B15AA392AB6CB CRC64;
MIKLLKPMSL SILLVFILSF SFPFPTAKAA ANDNNVEWNG LFHDQGPLFD NAPEPTSTQS
VTLKLRTFKG DITSANIKYW DTADNAFHWV PMVWDSNDPT GTFDYWKGTI PASPSIKYYR
FQINDGTSTA WYNGNGPSST EPNADDFYII PNFKTPDWLK NGVMYQIFPD RFYNGDSSND
VQTGSYTYNG TPTEKKAWGS SVYADPGYDN SLVFFGGDLA GIDQKLGYIK KTLGANILYL
NPIFKAPTNH KYDTQDYMAV DPAFGDNSTL QTLINDIHST ANGPKGYLIL DGVFNHTGDS
HPWFDKYNNF SSQGAYESQS SPWYNYYTFY TWPDSYASFL GFNSLPKLNY GNSGSAVRGV
IYNNSNSVAK TYLNPPYSVD GWRLDAAQYV DANGNNGSDV TNHQIWSEFR NAVKGVNSNA
AIIGEYWGNA NPWTAQGNQW DAATNFDGFT QPVSEWITGK DYQNNSASIS TTQFDSWLRG
TRANYPTNVQ QSMMNFLSNH DITRFATRSG GDLWKTYLAL IFQMTYVGTP TIYYGDEYGM
QGGADPDNRR SFDWSQATPS NSAVALTQKL ITIRNQYPAL RTGSFMTLIT DDTNKIYSYG
RFDNVNRIAV VLNNDSVSHT VNVPVWQLSM PNGSTVTDKI TGHSYTVQNG MVTVAVDGHY
GAVLAQ
