MPADE_PENRF
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};
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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
