PRX2_PENRF
ID PRX2_PENRF Reviewed; 342 AA.
AC W6Q4Q9;
DT 07-OCT-2020, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 28.
DE RecName: Full=Aristolochene synthase {ECO:0000303|PubMed:15186158};
DE Short=AS {ECO:0000303|PubMed:15186158};
DE EC=4.2.3.9 {ECO:0000269|PubMed:15186158, ECO:0000269|PubMed:8440737};
DE AltName: Full=PR-toxin biosynthesis cluster protein 2 {ECO:0000303|PubMed:24239699};
DE AltName: Full=Sesquiterpene cyclase {ECO:0000303|PubMed:15186158};
GN Name=prx2 {ECO:0000303|PubMed:24239699};
GN Synonyms=ari1 {ECO:0000303|PubMed:15186158},
GN ORF2 {ECO:0000303|PubMed:27921136}; ORFNames=PROQFM164_S02g001465;
OS Penicillium roqueforti (strain FM164).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=1365484;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=FM164;
RX PubMed=24407037; DOI=10.1038/ncomms3876;
RA Cheeseman K., Ropars J., Renault P., Dupont J., Gouzy J., Branca A.,
RA Abraham A.L., Ceppi M., Conseiller E., Debuchy R., Malagnac F., Goarin A.,
RA Silar P., Lacoste S., Sallet E., Bensimon A., Giraud T., Brygoo Y.;
RT "Multiple recent horizontal transfers of a large genomic region in cheese
RT making fungi.";
RL Nat. Commun. 5:2876-2876(2014).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8440737; DOI=10.1016/S0021-9258(18)53644-9;
RA Proctor R.H., Hohn T.M.;
RT "Aristolochene synthase. Isolation, characterization, and bacterial
RT expression of a sesquiterpenoid biosynthetic gene (Ari1) from Penicillium
RT roqueforti.";
RL J. Biol. Chem. 268:4543-4548(1993).
RN [3]
RP FUNCTION.
RX PubMed=16345540; DOI=10.1128/aem.39.4.770-776.1980;
RA Moreau S., Lablache-Combier A., Biguet J.;
RT "Production of eremofortins A, B, and C relative to formation of PR toxin
RT by Penicillium roqueforti.";
RL Appl. Environ. Microbiol. 39:770-776(1980).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF TYR-92; ASP-115; ASN-244; SER-248 AND GLU-252.
RX PubMed=15186158; DOI=10.1021/ja0499593;
RA Felicetti B., Cane D.E.;
RT "Aristolochene synthase: mechanistic analysis of active site residues by
RT site-directed mutagenesis.";
RL J. Am. Chem. Soc. 126:7212-7221(2004).
RN [5]
RP FUNCTION, DISRUPTION PHENOTYPE, AND PATHWAY.
RX PubMed=24239699; DOI=10.1016/j.fgb.2013.10.009;
RA Hidalgo P.I., Ullan R.V., Albillos S.M., Montero O., Fernandez-Bodega M.A.,
RA Garcia-Estrada C., Fernandez-Aguado M., Martin J.F.;
RT "Molecular characterization of the PR-toxin gene cluster in Penicillium
RT roqueforti and Penicillium chrysogenum: cross talk of secondary metabolite
RT pathways.";
RL Fungal Genet. Biol. 62:11-24(2014).
RN [6]
RP FUNCTION.
RX PubMed=26274339; DOI=10.1002/anie.201506128;
RA Riclea R., Dickschat J.S.;
RT "Identification of intermediates in the biosynthesis of PR toxin by
RT Penicillium roqueforti.";
RL Angew. Chem. Int. Ed. 54:12167-12170(2015).
RN [7]
RP FUNCTION, AND PATHWAY.
RX PubMed=27921136; DOI=10.1007/s00253-016-7995-5;
RA Hidalgo P.I., Poirier E., Ullan R.V., Piqueras J., Meslet-Cladiere L.,
RA Coton E., Coton M.;
RT "Penicillium roqueforti PR toxin gene cluster characterization.";
RL Appl. Microbiol. Biotechnol. 101:2043-2056(2017).
CC -!- FUNCTION: Aristolochene synthase; part of the gene cluster that
CC mediates the biosynthesis of PR-toxin, a bicyclic sesquiterpene
CC belonging to the eremophilane class and acting as a mycotoxin
CC (PubMed:24239699, PubMed:27921136). The first step of the pathway is
CC catalyzed by the aristolochene synthase which performs the cyclization
CC of trans,trans-farnesyl diphosphate (FPP) to the bicyclic sesquiterpene
CC aristolochene (PubMed:8440737, PubMed:15186158, PubMed:24239699).
CC Following the formation of aristolochene, the non-oxygenated
CC aristolochene is converted to the trioxygenated intermediate
CC eremofortin B, via 7-epi-neopetasone (PubMed:24239699,
CC PubMed:26274339). This conversion appears to involve three enzymes, a
CC hydroxysterol oxidase-like enzyme, the quinone-oxidase prx3 that forms
CC the quinone-type-structure in the bicyclic nucleus of aristolochene
CC with the C8-oxo group and the C-3 hydroxyl group, and the P450
CC monooxygenase ORF6 that introduces the epoxide at the double bond
CC between carbons 1 and 2 (PubMed:24239699, PubMed:27921136). No monoxy
CC or dioxy-intermediates have been reported to be released to the broth,
CC so these three early oxidative reactions may be coupled together
CC (PubMed:24239699). Eremofortin B is further oxidized by another P450
CC monooxygenase, that introduces a second epoxide between carbons 7 and
CC 11 prior to acetylation to eremofortin A by the acetyltransferase ORF8
CC (PubMed:16345540, PubMed:24239699, PubMed:27921136). The second
CC epoxidation may be performed by a second P450 monooxygenase
CC (PubMed:24239699). After the acetylation step, eremofortin A is
CC converted to eremofortin C and then to PR-toxin (PubMed:24239699).
CC First the conversion of eremofortin A to eremofortin C proceeds by
CC oxidation of the side chain of the molecule at C-12 and is catalyzed by
CC the short-chain oxidoreductase prx1 (PubMed:16345540, PubMed:24239699).
CC The cytochrome P450 monooxygenase ORF5 also plays a role in this step
CC (PubMed:27921136). The primary alcohol formed at C-12 is finally
CC oxidized by the short-chain alcohol dehydrogenase prx4 that forms PR-
CC toxin (PubMed:16345540, PubMed:24239699). {ECO:0000269|PubMed:15186158,
CC ECO:0000269|PubMed:16345540, ECO:0000269|PubMed:24239699,
CC ECO:0000269|PubMed:26274339, ECO:0000269|PubMed:27921136,
CC ECO:0000269|PubMed:8440737}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E)-farnesyl diphosphate = (+)-aristolochene +
CC diphosphate; Xref=Rhea:RHEA:19825, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:43445, ChEBI:CHEBI:175763; EC=4.2.3.9;
CC Evidence={ECO:0000269|PubMed:15186158, ECO:0000269|PubMed:8440737};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q9UR08};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250|UniProtKB:Q9UR08};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.6 uM for (2E,6E)-farnesyl diphosphate
CC {ECO:0000269|PubMed:15186158};
CC -!- PATHWAY: Sesquiterpene biosynthesis; aristolochene biosynthesis;
CC aristolochene from farnesyl diphosphate: step 1/1.
CC {ECO:0000269|PubMed:15186158, ECO:0000269|PubMed:8440737}.
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:Q9UR08}.
CC -!- DISRUPTION PHENOTYPE: Reduces the production of PR-toxin and leads to a
CC large increase in mycophenolic acid production.
CC {ECO:0000269|PubMed:24239699}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; HG792016; CDM31315.1; -; Genomic_DNA.
DR AlphaFoldDB; W6Q4Q9; -.
DR SMR; W6Q4Q9; -.
DR STRING; 1365484.W6Q4Q9; -.
DR EnsemblFungi; CDM31315; CDM31315; PROQFM164_S02g001465.
DR OrthoDB; 1143139at2759; -.
DR UniPathway; UPA00177; UER00582.
DR Proteomes; UP000030686; Unassembled WGS sequence.
DR GO; GO:0045483; F:aristolochene synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW Lyase; Magnesium; Metal-binding.
FT CHAIN 1..342
FT /note="Aristolochene synthase"
FT /id="PRO_0000451228"
FT BINDING 115
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 115
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 115
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT BINDING 200
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 244
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 244
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT BINDING 248
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 251
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 252
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT SITE 92
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT SITE 112
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT SITE 178
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT SITE 334
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:Q03471"
FT MUTAGEN 92
FT /note="Y->F: Causes 100-fold reduction in kcat but a 50-
FT fold decrease in KM, resulting in a 2-fold decrease in
FT catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:15186158"
FT MUTAGEN 115
FT /note="D->N: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:15186158"
FT MUTAGEN 244
FT /note="N->L: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:15186158"
FT MUTAGEN 248
FT /note="S->A: Abolishes catalytic activity; when associated
FT with D-252."
FT /evidence="ECO:0000269|PubMed:15186158"
FT MUTAGEN 252
FT /note="E->D: Abolishes catalytic activity; when associated
FT with A-248."
FT /evidence="ECO:0000269|PubMed:15186158"
SQ SEQUENCE 342 AA; 39192 MW; ACFBD63F4FF9BB03 CRC64;
MATSTETISS LAQPFVHLEN PINSPLVKET IRPRNDTTIT PPPTQWSYLC HPRVKEVQDE
VDGYFLENWK FPSFKAVRTF LDAKFSEVTC LYFPLALDDR IHFACRLLTV LFLIDDVLEH
MSFADGEAYN NRLIPISRGD VLPDRTKPEE FILYDLWESM RAHDAELANE VLEPTFVFMR
AQTDRARLSI HELGHYLEYR EKDVGKALLS ALMRFSMGLR LSADELQDMK ALEANCAKQL
SVVNDIYSYD KEEEASRTGH KEGAFLCSAV KVLAEESKLG IPATKRVLWS MTREWETVHD
EIVAEKIASP DGCSEAAKAY MKGLEYQMSG NEQWSKTTRR YN
