OTASE_ASPNC
ID OTASE_ASPNC Reviewed; 480 AA.
AC A2R2V4;
DT 29-SEP-2021, integrated into UniProtKB/Swiss-Prot.
DT 06-MAR-2007, sequence version 1.
DT 03-AUG-2022, entry version 75.
DE RecName: Full=Ochratoxinase {ECO:0000303|PubMed:24947135};
DE Short=OTase {ECO:0000303|PubMed:24947135};
DE EC=3.4.17.- {ECO:0000269|PubMed:24947135};
DE AltName: Full=Amidohydrolase 2 {ECO:0000303|PubMed:24947135};
DE Short=Amidase 2 {ECO:0000303|PubMed:24947135};
DE AltName: Full=Carboxypeptidase Am2 {ECO:0000303|PubMed:33647354};
GN Name=Am2 {ECO:0000303|PubMed:24947135}; ORFNames=An14g02080;
OS Aspergillus niger (strain CBS 513.88 / FGSC A1513).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=425011;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=CBS 513.88 / FGSC A1513 / ATCC MYA-4892;
RX PubMed=17259976; DOI=10.1038/nbt1282;
RA Pel H.J., de Winde J.H., Archer D.B., Dyer P.S., Hofmann G., Schaap P.J.,
RA Turner G., de Vries R.P., Albang R., Albermann K., Andersen M.R.,
RA Bendtsen J.D., Benen J.A.E., van den Berg M., Breestraat S., Caddick M.X.,
RA Contreras R., Cornell M., Coutinho P.M., Danchin E.G.J., Debets A.J.M.,
RA Dekker P., van Dijck P.W.M., van Dijk A., Dijkhuizen L., Driessen A.J.M.,
RA d'Enfert C., Geysens S., Goosen C., Groot G.S.P., de Groot P.W.J.,
RA Guillemette T., Henrissat B., Herweijer M., van den Hombergh J.P.T.W.,
RA van den Hondel C.A.M.J.J., van der Heijden R.T.J.M., van der Kaaij R.M.,
RA Klis F.M., Kools H.J., Kubicek C.P., van Kuyk P.A., Lauber J., Lu X.,
RA van der Maarel M.J.E.C., Meulenberg R., Menke H., Mortimer M.A.,
RA Nielsen J., Oliver S.G., Olsthoorn M., Pal K., van Peij N.N.M.E.,
RA Ram A.F.J., Rinas U., Roubos J.A., Sagt C.M.J., Schmoll M., Sun J.,
RA Ussery D., Varga J., Vervecken W., van de Vondervoort P.J.J., Wedler H.,
RA Woesten H.A.B., Zeng A.-P., van Ooyen A.J.J., Visser J., Stam H.;
RT "Genome sequencing and analysis of the versatile cell factory Aspergillus
RT niger CBS 513.88.";
RL Nat. Biotechnol. 25:221-231(2007).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF SER-135; TYR-160 AND
RP TYR-206.
RX PubMed=33647354; DOI=10.1016/j.jbiotec.2021.02.015;
RA Xiong K., Liu J., Wang X., Sun B., Zhang Y., Zhao Z., Pei P., Li X.;
RT "Engineering a carboxypeptidase from Aspergillus niger M00988 by mutation
RT to increase its ability in high Fischer ratio oligopeptide preparation.";
RL J. Biotechnol. 330:1-8(2021).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) OF 43-480 IN COMPLEX WITH ZINC,
RP FUNCTION, SUBCELLULAR LOCATION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, DOMAIN, COFACTOR, ACTIVE SITE, ACTIVITY REGULATION,
RP AND BIOTECHNOLOGY.
RX PubMed=24947135; DOI=10.1042/bj20140382;
RA Dobritzsch D., Wang H., Schneider G., Yu S.;
RT "Structural and functional characterization of ochratoxinase, a novel
RT mycotoxin-degrading enzyme.";
RL Biochem. J. 462:441-452(2014).
CC -!- FUNCTION: Carboxypeptidase that catalyzes the release of a C-terminal
CC amino acid with specific catalytic activity for aromatic amino acids
CC such as phenylalanine (PubMed:24947135, PubMed:33647354). Is able to
CC degrade ochratoxin A, one of the five major mycotoxins most harmful to
CC humans and animals that is produced by Aspergillus and Penicillium
CC species and occurs in a wide range of agricultural products
CC (PubMed:24947135). {ECO:0000269|PubMed:24947135,
CC ECO:0000269|PubMed:33647354}.
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:24947135};
CC -!- ACTIVITY REGULATION: The Zn(2+)-specific chelator 1,10-phenanthroline
CC inhibits the enzyme activity. {ECO:0000269|PubMed:24947135}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 6. {ECO:0000269|PubMed:24947135};
CC Temperature dependence:
CC Optimum temperature is 66 degrees Celsius.
CC {ECO:0000269|PubMed:24947135};
CC -!- SUBUNIT: Homooctamer. {ECO:0000269|PubMed:24947135}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:24947135}.
CC -!- DOMAIN: Each subunit of the homooctomeric enzyme folds into a two-
CC domain structure characteristic of a metal dependent amidohydrolase,
CC with a twisted TIM (triosephosphateisomerase)-barrel and a smaller
CC beta-sandwich domain. {ECO:0000269|PubMed:24947135}.
CC -!- BIOTECHNOLOGY: Detoxification of contaminated food is a challenging
CC health issue and ochratoxinase is a promising enzyme that hydrolyzes
CC ochrtoxin A, a mycotoxin demonstrated to have nephrotoxic, immunotoxic,
CC genotoxic, neurotoxic, and teratogenic properties; and that occurs in a
CC wide range of agricultural products (PubMed:24947135). Its
CC carboxypeptidase activity allows also to produce high Fischer ratio
CC (the ratio of moles of branched chain amino acid content to aromatic
CC amino acid content) oligopeptides, a kind of functional oligopeptides
CC that protect the liver, treat phenylketonuria, have anti-fatigue
CC properties, exhibits antioxidant properties, and support
CC anticoagulation of the blood (PubMed:33647354). Producing high Fischer
CC ratio oligopeptides using an enzymatic method is therefore of great
CC health significance and economic benefit (PubMed:33647354).
CC {ECO:0000269|PubMed:24947135, ECO:0000269|PubMed:33647354}.
CC -!- SIMILARITY: Belongs to the metallo-dependent hydrolases superfamily.
CC Ochratoxinase amidase 2 family. {ECO:0000305}.
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=Small poison - Issue 243 of
CC January 2022;
CC URL="https://web.expasy.org/spotlight/back_issues/243/";
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DR EMBL; AM270317; CAK41945.1; -; Genomic_DNA.
DR PDB; 4C5Y; X-ray; 3.00 A; A/B=43-480.
DR PDB; 4C60; X-ray; 2.50 A; A/B/C/D/E/F/G/H=43-480.
DR PDB; 4C65; X-ray; 2.20 A; A/B/C/D/E/F/G/H=43-480.
DR PDBsum; 4C5Y; -.
DR PDBsum; 4C60; -.
DR PDBsum; 4C65; -.
DR SMR; A2R2V4; -.
DR PaxDb; A2R2V4; -.
DR EnsemblFungi; CAK41945; CAK41945; An14g02080.
DR VEuPathDB; FungiDB:An14g02080; -.
DR HOGENOM; CLU_023620_2_1_1; -.
DR Proteomes; UP000006706; Chromosome 1R.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004180; F:carboxypeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0016810; F:hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008237; F:metallopeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:1900817; P:ochratoxin A catabolic process; IDA:CACAO.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR InterPro; IPR006680; Amidohydro-rel.
DR InterPro; IPR011059; Metal-dep_hydrolase_composite.
DR InterPro; IPR032466; Metal_Hydrolase.
DR Pfam; PF01979; Amidohydro_1; 1.
DR SUPFAM; SSF51338; SSF51338; 2.
DR SUPFAM; SSF51556; SSF51556; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carboxypeptidase; Hydrolase; Metal-binding; Metalloprotease;
KW Protease; Reference proteome; Secreted; Zinc.
FT CHAIN 1..480
FT /note="Ochratoxinase"
FT /id="PRO_0000453663"
FT ACT_SITE 246
FT /evidence="ECO:0000269|PubMed:24947135"
FT ACT_SITE 378
FT /evidence="ECO:0000269|PubMed:24947135"
FT BINDING 111
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT BINDING 113
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT BINDING 246
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT BINDING 246
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT BINDING 287
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT BINDING 307
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24947135,
FT ECO:0007744|PDB:4C5Y"
FT MUTAGEN 135
FT /note="S->G,W: Affect substrate binding and
FT carboxypeptidase activity."
FT /evidence="ECO:0000269|PubMed:33647354"
FT MUTAGEN 160
FT /note="Y->S,G: Affect substrate binding and
FT carboxypeptidase activity."
FT /evidence="ECO:0000269|PubMed:33647354"
FT MUTAGEN 206
FT /note="Y->S,G: Affect substrate binding and
FT carboxypeptidase activity."
FT /evidence="ECO:0000269|PubMed:33647354"
FT STRAND 49..57
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 60..62
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 65..73
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 76..82
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 83..85
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 88..93
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 95..105
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 107..112
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 125..129
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 132..148
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 151..156
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 158..160
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 161..169
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 177..180
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 182..186
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 199..206
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 207..210
FT /evidence="ECO:0007829|PDB:4C65"
FT TURN 213..217
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 219..223
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 227..239
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 243..248
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 256..258
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 267..279
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 284..288
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 291..300
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 304..307
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 313..322
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 325..327
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 330..339
FT /evidence="ECO:0007829|PDB:4C65"
FT TURN 340..343
FT /evidence="ECO:0007829|PDB:4C60"
FT HELIX 346..369
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 386..394
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 400..407
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 411..418
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 433..438
FT /evidence="ECO:0007829|PDB:4C65"
FT TURN 440..442
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 444..448
FT /evidence="ECO:0007829|PDB:4C65"
FT HELIX 450..452
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 453..458
FT /evidence="ECO:0007829|PDB:4C65"
FT STRAND 461..465
FT /evidence="ECO:0007829|PDB:4C65"
SQ SEQUENCE 480 AA; 51200 MW; 38F48AD24CCA5030 CRC64;
MVRRIASATP MVQSPMSPLG TTYCVRPNPV SLNLQRRPLV IASTDEAKVT IIYAGLLIPG
DGEPLRNAAL VISDKIIAFV GSEADIPKKY LRSTQSTHRV PVLMPGLWDC HMHFGGDDDY
YNDYTSGLAT HPASSGARLA RGCWEALQNG YTSYRDLAGY GCEVAKAIND GTIVGPNVYS
SGAALSQTAG HGDIFALPAG EVLGSYGVMN PRPGYWGAGP LCIADGVEEV RRAVRLQIRR
GAKVIKVMAS GGVMSRDDNP NFAQFSPEEL KVIVEEAARQ NRIVSAHVHG KAGIMAAIKA
GCKSLEHVSY ADEEVWELMK EKGILYVATR SVIEIFLASN GEGLVKESWA KLQALADSHL
KAYQGAIKAG VTIALGTDTA PGGPTALELQ FAVERGGMTP LEAIKAATAN APLSVGPQAP
LTGQLREGYE ADVIALEENP LEDIKVFQEP KAVTHVWKGG KLFKGPGIGP WGEDARNPFL
