ID MPADE_PENRF Reviewed; 852 AA. AC W6QRN8; DT 26-FEB-2020, integrated into UniProtKB/Swiss-Prot. DT 10-FEB-2021, sequence version 2. DT 03-AUG-2022, entry version 35. DE RecName: Full=Cytochrome P450 monooxygenase mpaDE {ECO:0000303|PubMed:26751579}; DE EC=1.-.-.- {ECO:0000305|PubMed:26751579}; DE AltName: Full=Mycophenolic acid biosynthesis cluster fusion protein DE {ECO:0000303|PubMed:26751579}; GN Name=mpaDE {ECO:0000303|PubMed:26751579}; ORFNames=PROQFM164_S05g000557; 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, DISRUPTION PHENOTYPE, AND PATHWAY. RX PubMed=26751579; DOI=10.1371/journal.pone.0147047; RA Del-Cid A., Gil-Duran C., Vaca I., Rojas-Aedo J.F., Garcia-Rico R.O., RA Levican G., Chavez R.; RT "Identification and functional analysis of the mycophenolic acid gene RT cluster of Penicillium roqueforti."; RL PLoS ONE 11:E0147047-E0147047(2016). CC -!- FUNCTION: Cytochrome P450 monooxygenase; part of the gene cluster that CC mediates the biosynthesis of mycophenolic acid (MPA), the first CC isolated antibiotic natural product in the world obtained from a CC culture of Penicillium brevicompactum in 1893 (PubMed:26751579). MpaDE CC is an endoplasmic reticulum-bound entyme that catalyzes the conversion CC of 5-methylorsellinic acid (5MOA) into the phthalide compound 5,7- CC dihydroxy-4,6-dimethylphthalide (DHMP) (PubMed:26751579). MpaDE first CC catalyzes hydroxylation of 5-MOA to 4,6-dihydroxy-2-(hydroxymethyl)-3- CC methylbenzoic acid (DHMB), and then acts as a lactone synthase that CC catalyzes the ring closure to convert DHMB into DHMP (PubMed:26751579). CC The first step of the pathway is the synthesis of 5-methylorsellinic CC acid (5MOA) by the cytosolic polyketide synthase mpaC. 5MOA is then CC converted to the phthalide compound 5,7-dihydroxy-4,6-dimethylphthalide CC (DHMP) by the endoplasmic reticulum-bound cytochrome P450 monooxygenase CC mpaDE. MpaDE first catalyzes hydroxylation of 5-MOA to 4,6-dihydroxy-2- CC (hydroxymethyl)-3-methylbenzoic acid (DHMB). MpaDE then acts as a CC lactone synthase that catalyzes the ring closure to convert DHMB into CC DHMP. The next step is the prenylation of DHMP by the Golgi apparatus- CC associated prenyltransferase mpaA to yield farnesyl-DHMP (FDHMP). The CC ER-bound oxygenase mpaB then mediates the oxidative cleavage the C19- CC C20 double bond in FDHMP to yield FDHMP-3C via a mycophenolic aldehyde CC intermediate. The O-methyltransferase mpaG catalyzes the methylation of CC FDHMP-3C to yield MFDHMP-3C. After the cytosolic methylation of FDHMP- CC 3C, MFDHMP-3C enters into peroxisomes probably via free diffusion due CC to its low molecular weight. Upon a peroxisomal CoA ligation reaction, CC catalyzed by a beta-oxidation component enzyme acyl-CoA ligase ACL891, CC MFDHMP-3C-CoA would then be restricted to peroxisomes for the following CC beta-oxidation pathway steps. The peroxisomal beta-oxidation machinery CC than converts MFDHMP-3C-CoA into MPA_CoA, via a beta-oxidation chain- CC shortening process. Finally mpaH acts as a peroxisomal acyl-CoA CC hydrolase with high substrate specificity toward MPA-CoA to release the CC final product MPA (PubMed:26751579) (Probable). CC {ECO:0000269|PubMed:26751579, ECO:0000305|PubMed:26751579}. CC -!- CATALYTIC ACTIVITY: CC Reaction=5-methylorsellinate + O2 + reduced [NADPH--hemoprotein CC reductase] = 4,6-dihydroxy-2-(hydroxymethyl)-3-methylbenzoate + H(+) CC + H2O + oxidized [NADPH--hemoprotein reductase]; CC Xref=Rhea:RHEA:66668, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965, CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210, ChEBI:CHEBI:146172, CC ChEBI:CHEBI:167385; Evidence={ECO:0000305|PubMed:26751579}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66669; CC Evidence={ECO:0000305|PubMed:26751579}; CC -!- CATALYTIC ACTIVITY: CC Reaction=4,6-dihydroxy-2-(hydroxymethyl)-3-methylbenzoate + H(+) = 5,7- CC dihydroxy-4-methylphthalide + H2O; Xref=Rhea:RHEA:66672, CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:68194, CC ChEBI:CHEBI:167385; Evidence={ECO:0000305|PubMed:26751579}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66673; CC Evidence={ECO:0000305|PubMed:26751579}; CC -!- COFACTOR: CC Name=heme; Xref=ChEBI:CHEBI:30413; CC Evidence={ECO:0000250|UniProtKB:P04798}; CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis. CC {ECO:0000269|PubMed:26751579}. CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane CC {ECO:0000250|UniProtKB:A0A0B5KYT4}; Single-pass membrane protein CC {ECO:0000255}. CC -!- DISRUPTION PHENOTYPE: Results in dramatic reduction in MPA production CC and leads to the accumulation of 5-MOA. {ECO:0000269|PubMed:26751579}. CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}. CC -!- SEQUENCE CAUTION: CC Sequence=CDM36724.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={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; HG792019; CDM36724.1; ALT_INIT; Genomic_DNA. DR AlphaFoldDB; W6QRN8; -. DR SMR; W6QRN8; -. DR EnsemblFungi; CDM36724; CDM36724; PROQFM164_S05g000557. DR OrthoDB; 467733at2759; -. DR UniPathway; UPA00213; -. DR Proteomes; UP000030686; Unassembled WGS sequence. DR GO; GO:0005789; C:endoplasmic reticulum membrane; ISS:GO_Central. DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW. DR GO; GO:0020037; F:heme binding; IEA:InterPro. DR GO; GO:0005506; F:iron ion binding; IEA:InterPro. DR GO; GO:0004497; F:monooxygenase activity; IMP:GO_Central. DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro. DR GO; GO:0140722; P:mycophenolic acid biosynthetic process; IMP:GO_Central. DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway. DR Gene3D; 1.10.630.10; -; 1. DR Gene3D; 3.60.15.10; -; 1. DR InterPro; IPR001128; Cyt_P450. DR InterPro; IPR017972; Cyt_P450_CS. DR InterPro; IPR002403; Cyt_P450_E_grp-IV. DR InterPro; IPR036396; Cyt_P450_sf. DR InterPro; IPR001279; Metallo-B-lactamas. DR InterPro; IPR036866; RibonucZ/Hydroxyglut_hydro. DR Pfam; PF00753; Lactamase_B; 1. DR Pfam; PF00067; p450; 1. DR PRINTS; PR00465; EP450IV. DR PRINTS; PR00385; P450. DR SMART; SM00849; Lactamase_B; 1. DR SUPFAM; SSF48264; SSF48264; 1. DR SUPFAM; SSF56281; SSF56281; 1. DR PROSITE; PS00086; CYTOCHROME_P450; 1. PE 3: Inferred from homology; KW Endoplasmic reticulum; Heme; Iron; Membrane; Metal-binding; Monooxygenase; KW Oxidoreductase; Transmembrane; Transmembrane helix. FT CHAIN 1..852 FT /note="Cytochrome P450 monooxygenase mpaDE" FT /id="PRO_0000449216" FT TOPO_DOM 1..6 FT /note="Lumenal" FT /evidence="ECO:0000305" FT TRANSMEM 7..29 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 30..852 FT /note="Cytoplasmic" FT /evidence="ECO:0000305" FT BINDING 448 FT /ligand="heme" FT /ligand_id="ChEBI:CHEBI:30413" FT /ligand_part="Fe" FT /ligand_part_id="ChEBI:CHEBI:18248" FT /note="axial binding residue" FT /evidence="ECO:0000250|UniProtKB:P04798" SQ SEQUENCE 852 AA; 96209 MW; C48E72CDD5344FA2 CRC64; MDYLIIIRIT AVAVVLYLTR YVCCLYLHLQ DVPGPLFAKF TNLQRVWWVK SGRAHEYHRR MHAVYGPAVR FGPNMVSISD PRTIPAIYPS RPGFPKSDFY RTQKPYTPNK GAMPAVFNSQ DEDLHKRLRS PIAPLYSMTN VVKLESFVDQ TLAVLLEQLD GRFLGSNDVP FDLGSWLQYF AFDSMGTLTF SRRYGFLEQG RDMNGILGEI WKFMKRVSVM GQIPWFDEFC NTNPFIALFR SPTGFGVLKV VDKFILQRLA PREKDEVSDE KDMLSQFLNI QASNPDVMPW APRAWTFSNI MAGSDSTANV MRTIMYNLLV HRDTLSRLQD ELLESESSNG LSRTCPSWEK VRDLPYLDAC VLEALRLHPP FCLPFERVVP GGGLTVCETY LPAGTIVGIS PYMANRDKET FGNDADEWRP ERWLGLSHED RKRLENSLLT FGAGRRTCLG KNIAILEIKK LIPVLLLNYD IQIVNPENYK TENAWFFKQT GLQAVIRKRA KMERGSSNKD KPTLPPVLNI PPSSSTVDVR VIDPGTLLDL RPDLFWQPEL PGLRKVTAPT YCFLISVGTR HVLFDLGVRQ DWERLPPSVV AMIKSQTTIQ NPRNISDILD SDASSLGIRS TDIEAIIWSH AHFDHIGDPS TFPLSTELVV GPGIRDSHWP GFPTNPDAIN LNSDIQGRKV REISFERTEK EAIKIGSFDA LDYFGDGSFY LLNAAGHSIG HIGALARVTT SPDSFVFMGG DSCHHAGVLR PSKYLPCPSH SRHIPLSSES ESVFTLSPVL PSDYDAALKT VDNIKELDAY DNVFLILAHD STLKGNMDFY PLTINDWKAK GYGKQTKWLF YKDLEDAMEG TK //