INUE_ASPAW
ID INUE_ASPAW Reviewed; 537 AA.
AC Q96TU3;
DT 09-JUL-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 03-AUG-2022, entry version 81.
DE RecName: Full=Extracellular exo-inulinase inuE;
DE EC=3.2.1.80;
DE Flags: Precursor;
GN Name=inuE; Synonyms=inu1;
OS Aspergillus awamori (Black koji mold).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus.
OX NCBI_TaxID=105351;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 20-59 AND 128-139,
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=11829749; DOI=10.1042/0264-6021:3620131;
RA Arand M., Golubev A.M., Neto J.R., Polikarpov I., Wattiez R.,
RA Korneeva O.S., Eneyskaya E.V., Kulminskaya A.A., Shabalin K.A.,
RA Shishliannikov S.M., Chepurnaya O.V., Neustroev K.N.;
RT "Purification, characterization, gene cloning and preliminary X-ray data of
RT the exo-inulinase from Aspergillus awamori.";
RL Biochem. J. 362:131-135(2002).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.55 ANGSTROMS) OF 20-537 IN COMPLEX WITH FRUCTOSE,
RP ACTIVE SITE, AND GLYCOSYLATION AT ASN-67; ASN-111; ASN-300; ASN-398 AND
RP ASN-430.
RX PubMed=15522299; DOI=10.1016/j.jmb.2004.09.024;
RA Nagem R.A., Rojas A.L., Golubev A.M., Korneeva O.S., Eneyskaya E.V.,
RA Kulminskaya A.A., Neustroev K.N., Polikarpov I.;
RT "Crystal structure of exo-inulinase from Aspergillus awamori: the enzyme
RT fold and structural determinants of substrate recognition.";
RL J. Mol. Biol. 344:471-480(2004).
CC -!- FUNCTION: Exo-inulinase involved in utilization of the plant storage
CC polymer inulin, consisting of fructooligosaccharides with a degree of
CC polymerization (DP) value from 2 to 60. Splits off terminal fructose
CC units successively from the non-reducing end of the inulin molecule,
CC and also hydrolyzes levan, stachyose and raffinose.
CC {ECO:0000269|PubMed:11829749}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal, non-reducing (2->1)- and (2->6)-linked
CC beta-D-fructofuranose residues in fructans.; EC=3.2.1.80;
CC Evidence={ECO:0000269|PubMed:11829749};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.003 mM for inulin {ECO:0000269|PubMed:11829749};
CC KM=10.1 mM for stachyose {ECO:0000269|PubMed:11829749};
CC KM=8 mM for raffinose {ECO:0000269|PubMed:11829749};
CC Vmax=175 umol/min/mg enzyme toward inulin
CC {ECO:0000269|PubMed:11829749};
CC Vmax=1.2 umol/min/mg enzyme toward levan
CC {ECO:0000269|PubMed:11829749};
CC Vmax=80 umol/min/mg enzyme toward stachyose
CC {ECO:0000269|PubMed:11829749};
CC Vmax=120 umol/min/mg enzyme toward raffinose
CC {ECO:0000269|PubMed:11829749};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250}.
CC -!- INDUCTION: Expression is induced in presence of maltose, sucrose or
CC inulin and controlled by the catabolite repressor creA and by the
CC inulinolytic genes regulator inuR.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 32 family. {ECO:0000305}.
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DR EMBL; AJ315793; CAC44220.1; -; Genomic_DNA.
DR PDB; 1Y4W; X-ray; 1.55 A; A=20-537.
DR PDB; 1Y9G; X-ray; 1.87 A; A=20-537.
DR PDB; 1Y9M; X-ray; 1.89 A; A=20-537.
DR PDBsum; 1Y4W; -.
DR PDBsum; 1Y9G; -.
DR PDBsum; 1Y9M; -.
DR AlphaFoldDB; Q96TU3; -.
DR SMR; Q96TU3; -.
DR CAZy; GH32; Glycoside Hydrolase Family 32.
DR CLAE; INX32A_ASPAW; -.
DR iPTMnet; Q96TU3; -.
DR BRENDA; 3.2.1.80; 494.
DR EvolutionaryTrace; Q96TU3; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0051669; F:fructan beta-fructosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 2.115.10.20; -; 1.
DR InterPro; IPR013320; ConA-like_dom_sf.
DR InterPro; IPR001362; Glyco_hydro_32.
DR InterPro; IPR018053; Glyco_hydro_32_AS.
DR InterPro; IPR013189; Glyco_hydro_32_C.
DR InterPro; IPR013148; Glyco_hydro_32_N.
DR InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR Pfam; PF08244; Glyco_hydro_32C; 1.
DR Pfam; PF00251; Glyco_hydro_32N; 1.
DR SMART; SM00640; Glyco_32; 1.
DR SUPFAM; SSF49899; SSF49899; 1.
DR SUPFAM; SSF75005; SSF75005; 1.
DR PROSITE; PS00609; GLYCOSYL_HYDROL_F32; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Direct protein sequencing;
KW Glycoprotein; Glycosidase; Hydrolase; Polysaccharide degradation; Secreted;
KW Signal.
FT SIGNAL 1..19
FT /evidence="ECO:0000269|PubMed:11829749"
FT CHAIN 20..537
FT /note="Extracellular exo-inulinase inuE"
FT /id="PRO_5000067603"
FT ACT_SITE 41
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10067"
FT ACT_SITE 241
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10067"
FT BINDING 40..41
FT /ligand="substrate"
FT BINDING 57
FT /ligand="substrate"
FT BINDING 65
FT /ligand="substrate"
FT BINDING 103
FT /ligand="substrate"
FT BINDING 188..189
FT /ligand="substrate"
FT BINDING 241
FT /ligand="substrate"
FT BINDING 335
FT /ligand="substrate"
FT CARBOHYD 49
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 67
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:15522299"
FT CARBOHYD 111
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:15522299"
FT CARBOHYD 112
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 254
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 300
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:15522299"
FT CARBOHYD 398
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:15522299"
FT CARBOHYD 430
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000269|PubMed:15522299"
FT STRAND 29..31
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 35..48
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 51..58
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 63..65
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 69..80
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 82..88
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 92..94
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 99..108
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 113..115
FT /evidence="ECO:0007829|PDB:1Y9G"
FT STRAND 119..121
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 124..134
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 148..158
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 167..169
FT /evidence="ECO:0007829|PDB:1Y4W"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 186..195
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 196..199
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 200..207
FT /evidence="ECO:0007829|PDB:1Y4W"
FT HELIX 208..210
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 212..223
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 225..230
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 237..250
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 251..254
FT /evidence="ECO:0007829|PDB:1Y9G"
FT STRAND 256..266
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 276..284
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 289..291
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 293..295
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 298..301
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 304..306
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 308..311
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 313..317
FT /evidence="ECO:0007829|PDB:1Y4W"
FT HELIX 323..325
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 327..331
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 335..337
FT /evidence="ECO:0007829|PDB:1Y4W"
FT HELIX 338..340
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 344..347
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 354..361
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 364..371
FT /evidence="ECO:0007829|PDB:1Y4W"
FT HELIX 375..377
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 384..392
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 394..396
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 404..413
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 417..427
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 431..433
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 435..440
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 441..444
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 445..449
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 460..462
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 465..469
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 476..485
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 488..493
FT /evidence="ECO:0007829|PDB:1Y4W"
FT TURN 494..497
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 498..503
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 512..520
FT /evidence="ECO:0007829|PDB:1Y4W"
FT STRAND 522..532
FT /evidence="ECO:0007829|PDB:1Y4W"
SQ SEQUENCE 537 AA; 59171 MW; 1575700AF21D070B CRC64;
MAPLSKALSV FMLMGITYAF NYDQPYRGQY HFSPQKNWMN DPNGLLYHNG TYHLFFQYNP
GGIEWGNISW GHAISEDLTH WEEKPVALLA RGFGSDVTEM YFSGSAVADV NNTSGFGKDG
KTPLVAMYTS YYPVAQTLPS GQTVQEDQQS QSIAYSLDDG LTWTTYDAAN PVIPNPPSPY
EAEYQNFRDP FVFWHDESQK WVVVTSIAEL HKLAIYTSDN LKDWKLVSEF GPYNAQGGVW
ECPGLVKLPL DSGNSTKWVI TSGLNPGGPP GTVGSGTQYF VGEFDGTTFT PDADTVYPGN
STANWMDWGP DFYAAAGYNG LSLNDHVHIG WMNNWQYGAN IPTYPWRSAM AIPRHMALKT
IGSKATLVQQ PQEAWSSISN KRPIYSRTFK TLSEGSTNTT TTGETFKVDL SFSAKSKAST
FAIALRASAN FTEQTLVGYD FAKQQIFLDR THSGDVSFDE TFASVYHGPL TPDSTGVVKL
SIFVDRSSVE VFGGQGETTL TAQIFPSSDA VHARLASTGG TTEDVRADIY KIASTWN
