PRX2_PENRO
ID PRX2_PENRO Reviewed; 342 AA.
AC Q03471;
DT 01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1993, sequence version 1.
DT 03-AUG-2022, entry version 113.
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 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};
OS Penicillium roqueforti.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=5082;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 56-70; 76-84;
RP 245-251 AND 323-336, FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=ATCC 10110 / BCRC 32359 / CBS 221.30 / JCM 22842 / NBRC 5459 / NCTC
RC 588 / NRRL 849;
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 [2]
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 [3]
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 [4]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
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 [5]
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 [6]
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).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH SUBSTRATE.
RX PubMed=10825154; DOI=10.1074/jbc.m000433200;
RA Caruthers J.M., Kang I., Rynkiewicz M.J., Cane D.E., Christianson D.W.;
RT "Crystal structure determination of aristolochene synthase from the blue
RT cheese mold, Penicillium roqueforti.";
RL J. Biol. Chem. 275:25533-25539(2000).
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, the conversion of
CC eremofortin A to eremofortin C and then to PR-toxin requires only two
CC enzymes (PubMed:24239699). First the conversion of eremofortin A to
CC eremofortin C proceeds by oxidation of the side chain of the molecule
CC at C-12 and is catalyzed by the short-chain oxidoreductase prx1
CC (PubMed:16345540, PubMed:24239699). The cytochrome P450 monooxygenase
CC ORF5 also plays a role in this step (PubMed:27921136). The primary
CC alcohol formed at C-12 is finally oxidized by the short-chain alcohol
CC dehydrogenase prx4 that forms PR-toxin (PubMed:16345540,
CC 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 (PubMed:24239699).
CC {ECO:0000269|PubMed:24239699}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; L05193; AAA33694.1; -; Genomic_DNA.
DR PIR; A45462; A45462.
DR PDB; 1DGP; X-ray; 2.80 A; A/B=40-339.
DR PDB; 1DI1; X-ray; 2.50 A; A/B=40-339.
DR PDBsum; 1DGP; -.
DR PDBsum; 1DI1; -.
DR AlphaFoldDB; Q03471; -.
DR SMR; Q03471; -.
DR KEGG; ag:AAA33694; -.
DR PhylomeDB; Q03471; -.
DR BioCyc; MetaCyc:MON-16547; -.
DR BRENDA; 4.2.3.9; 4638.
DR SABIO-RK; Q03471; -.
DR UniPathway; UPA00177; UER00582.
DR EvolutionaryTrace; Q03471; -.
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 3D-structure; Direct protein sequencing; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..342
FT /note="Aristolochene synthase"
FT /id="PRO_0000064677"
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:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DGP"
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:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DGP"
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:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DI1"
FT SITE 112
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DI1"
FT SITE 178
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DI1"
FT SITE 334
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000269|PubMed:10825154,
FT ECO:0007744|PDB:1DI1"
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"
FT HELIX 54..68
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 74..83
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 85..92
FT /evidence="ECO:0007829|PDB:1DI1"
FT TURN 98..100
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 101..120
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 123..137
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 148..180
FT /evidence="ECO:0007829|PDB:1DI1"
FT TURN 181..183
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 193..201
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 204..217
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 223..227
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 230..248
FT /evidence="ECO:0007829|PDB:1DI1"
FT TURN 249..252
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 269..277
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 281..307
FT /evidence="ECO:0007829|PDB:1DI1"
FT HELIX 315..336
FT /evidence="ECO:0007829|PDB:1DI1"
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
