INUA_ASPNG
ID INUA_ASPNG Reviewed; 516 AA.
AC O74641; Q96WZ8;
DT 11-JUN-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1998, sequence version 1.
DT 03-AUG-2022, entry version 80.
DE RecName: Full=Extracellular endo-inulinase inuA;
DE EC=3.2.1.7;
DE Flags: Precursor;
GN Name=inuA;
OS Aspergillus niger.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=5061;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=12;
RX PubMed=9805373; DOI=10.1271/bbb.62.1731;
RA Ohta K., Akimoto H., Matsuda S., Toshimitsu D., Nakamura T.;
RT "Molecular cloning and sequence analysis of two endoinulinase genes from
RT Aspergillus niger.";
RL Biosci. Biotechnol. Biochem. 62:1731-1738(1998).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=12;
RX PubMed=16232670; DOI=10.1016/s1389-1723(00)87086-3;
RA Akimoto H., Kushima T., Nakamura T., Ohta K.;
RT "Transcriptional analysis of two endoinulinase genes inuA and inuB in
RT Aspergillus niger and nucleotide sequences of their promoter regions.";
RL J. Biosci. Bioeng. 88:599-604(1999).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], VARIANTS THR-18; ALA-110; VAL-130;
RP ASN-292; GLU-371; LYS-419; THR-437 AND ASP-455, FUNCTION, SUBCELLULAR
RP LOCATION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY
RP REGULATION.
RC STRAIN=AF10;
RX PubMed=15135402; DOI=10.1016/j.pep.2004.02.015;
RA Zhang L., Zhao C., Zhu D., Ohta Y., Wang Y.;
RT "Purification and characterization of inulinase from Aspergillus niger AF10
RT expressed in Pichia pastoris.";
RL Protein Expr. Purif. 35:272-275(2004).
RN [4]
RP BIOTECHNOLOGY.
RX PubMed=8481000; DOI=10.1128/aem.59.3.729-733.1993;
RA Ohta K., Hamada S., Nakamura T.;
RT "Production of high concentrations of ethanol from inulin by simultaneous
RT saccharification and fermentation using Aspergillus niger and Saccharomyces
RT cerevisiae.";
RL Appl. Environ. Microbiol. 59:729-733(1993).
CC -!- FUNCTION: Endo-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. {ECO:0000269|PubMed:15135402}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endohydrolysis of (2->1)-beta-D-fructosidic linkages in
CC inulin.; EC=3.2.1.7; Evidence={ECO:0000269|PubMed:15135402};
CC -!- ACTIVITY REGULATION: Activity is stimulated by Mn(2+), Fe(2+) Ca(2+)
CC metal ions and DTT; and inhibited by glucose, Mg(2+), Zn(2+), Cu(2+),
CC Hg(2+), Al(3+), and Fe(3+). {ECO:0000269|PubMed:15135402}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.57 mM for inulin {ECO:0000269|PubMed:15135402};
CC pH dependence:
CC Optimum pH is 4.5. {ECO:0000269|PubMed:15135402};
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius.
CC {ECO:0000269|PubMed:15135402};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250}.
CC -!- BIOTECHNOLOGY: Plays an important role in biofuel production. Pure
CC nonhydrolyzed inulin can be directly converted to ethanol in a
CC simultaneous saccharification and fermentation process when combining
CC A.niger and S.cerevisiae. Endoinulinase can digest
CC fructooligosaccharides with high degree of polymerization (DP) values
CC (>20) into short molecules that may be readily hydrolyzed by
CC S.cerevisiae SUC2. Thus, introduction of an endoinulinase gene into
CC S.cerevisiae will improve its inulin utilization and ethanol
CC fermentation through collaboration between the heterologous
CC endoinulinase and the invertase SUC2. {ECO:0000269|PubMed:8481000}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 32 family. {ECO:0000305}.
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DR EMBL; AB012771; BAA33797.1; -; Genomic_DNA.
DR EMBL; AF369388; AAK43726.1; -; Genomic_DNA.
DR PIR; JE0301; JE0301.
DR AlphaFoldDB; O74641; -.
DR SMR; O74641; -.
DR STRING; 5061.CADANGAP00008504; -.
DR CAZy; GH32; Glycoside Hydrolase Family 32.
DR CLAE; INU32A_ASPNG; -.
DR VEuPathDB; FungiDB:An11g03200; -.
DR VEuPathDB; FungiDB:ASPNIDRAFT2_1095959; -.
DR VEuPathDB; FungiDB:ATCC64974_89460; -.
DR VEuPathDB; FungiDB:M747DRAFT_330865; -.
DR eggNOG; KOG0228; Eukaryota.
DR BRENDA; 3.2.1.7; 518.
DR BRENDA; 3.2.1.80; 518.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0051670; F:inulinase 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; 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.
PE 1: Evidence at protein level;
KW Carbohydrate metabolism; Glycoprotein; Glycosidase; Hydrolase;
KW Polysaccharide degradation; Secreted; Signal.
FT SIGNAL 1..25
FT /evidence="ECO:0000255"
FT CHAIN 26..516
FT /note="Extracellular endo-inulinase inuA"
FT /id="PRO_5000049217"
FT ACT_SITE 43
FT /evidence="ECO:0000250"
FT BINDING 40..43
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 59
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 67
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 99..100
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 175..176
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 233
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT CARBOHYD 109
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 210
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 372
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 419
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT VARIANT 18
FT /note="M -> T (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 110
FT /note="T -> A (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 130
FT /note="T -> V (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 292
FT /note="D -> N (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 371
FT /note="A -> E (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 419
FT /note="N -> K (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 437
FT /note="I -> T (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
FT VARIANT 455
FT /note="G -> D (in strain: AF10)"
FT /evidence="ECO:0000269|PubMed:15135402"
SQ SEQUENCE 516 AA; 55797 MW; 85F64BF3701E7A37 CRC64;
MLNPKVAYMV WMTCLGLMLP SQAQSNDYRP SYHFTPDQYW MNEPNGLIKI GSTWHLFFQH
NPTANVWGNI CWGHATSTDL MHWAHKPTAI ADENGVEAFT GTAYYDPNNT SGLGDSANPP
YLAWFTGYTT SSQTQDQRLA FSVDNGATWT KFQGNPIIST SQEAPHDITG GLESRDPKVF
FHRQSGNWIM VLAHGGQDKL SFWTSADTIN WTWQSDLKST SINGLSSDIT GWEVPDMFEL
PVEGTEETTW VVMMTPAEGS PAGGNGVLAI TGSFDGKSFT ADPVDASTMW LDNGRDFDGA
LSWVNVPASD GRRIIAAVMN SYGSNPPTTT WKGMLSFPRT LSLKKVGTQQ HFVQQPITEL
DTISTSLQTL ANQTITPGQT LLSSIRGTAL DVRVAFYPDA GSVLSLAVRK GASEQTVINY
TQSDATLSVD RTESGDISYD PAAGGVHTAK LEEDGTGLVS IRVLVDTCSV EVFGGQGEAV
ISDLIFPSDS SDGLALEVTG GNAVLQSVDV RSVSLE
