NEPU2_THEVU
ID NEPU2_THEVU Reviewed; 585 AA.
AC Q08751;
DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1995, sequence version 1.
DT 03-AUG-2022, entry version 110.
DE RecName: Full=Neopullulanase 2;
DE EC=3.2.1.135 {ECO:0000269|PubMed:7763540};
DE AltName: Full=Alpha-amylase II;
DE AltName: Full=TVA II;
GN Name=tvaII;
OS Thermoactinomyces vulgaris.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Thermoactinomycetaceae;
OC Thermoactinomyces.
OX NCBI_TaxID=2026;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CHARACTERIZATION, FUNCTION, AND
RP CATALYTIC ACTIVITY.
RC STRAIN=R-47;
RX PubMed=7763540; DOI=10.1271/bbb.57.395;
RA Tonozuka T., Ohtsuka M., Mogi S., Sakai H., Ohta T., Sakano Y.;
RT "A neopullulanase-type alpha-amylase gene from Thermoactinomyces vulgaris
RT R-47.";
RL Biosci. Biotechnol. Biochem. 57:395-401(1993).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS).
RC STRAIN=R-47;
RX PubMed=10222200; DOI=10.1006/jmbi.1999.2647;
RA Kamitori S., Kondo S., Okuyama K., Yokota T., Shimura Y., Tonozuka T.,
RA Sakano Y.;
RT "Crystal structure of Thermoactinomyces vulgaris R-47 alpha-amylase II
RT (TVAII) hydrolyzing cyclodextrins and pullulan at 2.6-A resolution.";
RL J. Mol. Biol. 287:907-921(1999).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH CALCIUM, AND
RP COFACTOR.
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: Hydrolyzes pullulan efficiently but only a small amount of
CC starch. Endohydrolysis of 1,4-alpha-glucosidic linkages in pullulan to
CC form panose. Cleaves also (1-6)-alpha-glucosidic linkages to form
CC maltotriose. {ECO:0000269|PubMed:7763540}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of pullulan to panose (6-alpha-D-glucosylmaltose).;
CC EC=3.2.1.135; Evidence={ECO:0000269|PubMed:7763540};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:12051850};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000269|PubMed:12051850};
CC -!- SUBUNIT: Monomer.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR EMBL; D13178; BAA02473.1; -; Genomic_DNA.
DR PIR; JC1486; JC1486.
DR PDB; 1BVZ; X-ray; 2.60 A; A/B=1-585.
DR PDB; 1G1Y; X-ray; 3.00 A; A/B=1-585.
DR PDB; 1JF5; X-ray; 3.20 A; A/B=1-585.
DR PDB; 1JF6; X-ray; 3.20 A; A/B=1-585.
DR PDB; 1JI2; X-ray; 2.30 A; A/B=1-585.
DR PDB; 1JIB; X-ray; 3.30 A; A/B=1-585.
DR PDB; 1JL8; X-ray; 3.20 A; A/B=1-585.
DR PDB; 1VB9; X-ray; 2.20 A; A/B=1-585.
DR PDB; 1VFM; X-ray; 2.90 A; A/B=1-585.
DR PDB; 1VFO; X-ray; 2.81 A; A/B=1-585.
DR PDB; 1VFU; X-ray; 3.10 A; A/B=1-585.
DR PDB; 1WZK; X-ray; 2.30 A; A/B=1-585.
DR PDB; 1WZL; X-ray; 2.00 A; A/B=1-585.
DR PDB; 1WZM; X-ray; 3.20 A; A/B=1-585.
DR PDB; 2D2O; X-ray; 2.10 A; A/B=1-585.
DR PDB; 3A6O; X-ray; 2.80 A; A/B=1-585.
DR PDBsum; 1BVZ; -.
DR PDBsum; 1G1Y; -.
DR PDBsum; 1JF5; -.
DR PDBsum; 1JF6; -.
DR PDBsum; 1JI2; -.
DR PDBsum; 1JIB; -.
DR PDBsum; 1JL8; -.
DR PDBsum; 1VB9; -.
DR PDBsum; 1VFM; -.
DR PDBsum; 1VFO; -.
DR PDBsum; 1VFU; -.
DR PDBsum; 1WZK; -.
DR PDBsum; 1WZL; -.
DR PDBsum; 1WZM; -.
DR PDBsum; 2D2O; -.
DR PDBsum; 3A6O; -.
DR AlphaFoldDB; Q08751; -.
DR SMR; Q08751; -.
DR DrugBank; DB02379; Beta-D-Glucose.
DR DrugBank; DB03995; Betadex.
DR DrugBank; DB02237; Maltotetraose.
DR CAZy; CBM34; Carbohydrate-Binding Module Family 34.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR PRIDE; Q08751; -.
DR EvolutionaryTrace; Q08751; -.
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 Gene3D; 3.90.400.10; -; 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 InterPro; IPR032091; Malt_amylase_C.
DR InterPro; IPR045857; O16G_dom_2.
DR Pfam; PF00128; Alpha-amylase; 1.
DR Pfam; PF02903; Alpha-amylase_N; 1.
DR Pfam; PF16657; Malt_amylase_C; 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; Glycosidase; Hydrolase;
KW Metal-binding.
FT CHAIN 1..585
FT /note="Neopullulanase 2"
FT /id="PRO_0000054311"
FT ACT_SITE 325
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT ACT_SITE 354
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 143
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 145
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 148
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 149
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 169
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 171
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12051850,
FT ECO:0007744|PDB:1JI2"
FT BINDING 244
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 323
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 420..421
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 465
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT BINDING 469
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P38940"
FT SITE 421
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250"
FT HELIX 3..5
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 12..14
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 22..30
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 31..33
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 35..42
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 48..50
FT /evidence="ECO:0007829|PDB:1VFU"
FT STRAND 53..56
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 58..62
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 64..74
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 76..78
FT /evidence="ECO:0007829|PDB:1VFU"
FT STRAND 80..87
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 89..91
FT /evidence="ECO:0007829|PDB:1BVZ"
FT STRAND 93..97
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 100..104
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 105..108
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 111..113
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 118..120
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 127..130
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 133..136
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 138..140
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 146..148
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 172..177
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 179..185
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 189..192
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 199..202
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 207..212
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 214..216
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 219..230
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 231..233
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 235..240
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 250..258
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 259..261
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 265..267
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 270..274
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 278..280
FT /evidence="ECO:0007829|PDB:1BVZ"
FT STRAND 285..289
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 294..296
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 301..316
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 321..324
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 327..329
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 332..345
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 350..353
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 360..362
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 364..367
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 369..372
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 374..384
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 391..402
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 407..410
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 414..417
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 425..428
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 429..431
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 433..445
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 446..453
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 456..458
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 467..469
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 476..478
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 481..496
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 498..502
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 504..511
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 512..515
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 516..523
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 526..533
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 535..537
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 539..544
FT /evidence="ECO:0007829|PDB:1WZL"
FT HELIX 547..549
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 551..555
FT /evidence="ECO:0007829|PDB:1WZL"
FT TURN 556..558
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 561..563
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 568..573
FT /evidence="ECO:0007829|PDB:1WZL"
FT STRAND 578..582
FT /evidence="ECO:0007829|PDB:1WZL"
SQ SEQUENCE 585 AA; 67467 MW; E311813A05A7791A CRC64;
MLLEAIFHEA KGSYAYPISE TQLRVRLRAK KGDVVRCEVL YADRYASPEE ELAHALAGKA
GSDERFDYFE ALLECSTKRV KYVFLLTGPQ GEAVYFGETG FSAERSKAGV FQYAYIHRSE
VFTTPEWAKE AVIYQIFPER FANGDPSNDP PGTEQWAKDA RPRHDSFYGG DLKGVIDRLP
YLEELGVTAL YFTPIFASPS HHKYDTADYL AIDPQFGDLP TFRRLVDEAH RRGIKIILDA
VFNHAGDQFF AFRDVLQKGE QSRYKDWFFI EDFPVSKTSR TNYETFAVQV PAMPKLRTEN
PEVKEYLFDV ARFWMEQGID GWRLDVANEV DHAFWREFRR LVKSLNPDAL IVGEIWHDAS
GWLMGDQFDS VMNYLFRESV IRFFATGEIH AERFDAELTR ARMLYPEQAA QGLWNLLDSH
DTERFLTSCG GNEAKFRLAV LFQMTYLGTP LIYYGDEIGM AGATDPDCRR PMIWEEKEQN
RGLFEFYKEL IRLRHRLASL TRGNVRSWHA DKQANLYAFV RTVQDQHVGV VLNNRGEKQT
VLLQVPESGG KTWLDCLTGE EVHGKQGQLK LTLRPYQGMI LWNGR
