MPDE2_PENBR
ID MPDE2_PENBR Reviewed; 853 AA.
AC A0A0B5KYT4;
DT 10-FEB-2021, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-2015, sequence version 1.
DT 03-AUG-2022, entry version 29.
DE RecName: Full=Cytochrome P450 monooxygenase mpaDE' {ECO:0000303|PubMed:25630520};
DE EC=1.-.-.- {ECO:0000269|PubMed:31209052};
DE AltName: Full=Mycophenolic acid biosynthesis cluster protein DE' {ECO:0000303|PubMed:25630520};
GN Name=mpaDE' {ECO:0000303|PubMed:25630520};
OS Penicillium brevicompactum.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=5074;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NRRL864;
RX PubMed=25630520; DOI=10.1002/cbic.201402600;
RA Zhang W., Cao S., Qiu L., Qi F., Li Z., Yang Y., Huang S., Bai F., Liu C.,
RA Wan X., Li S.;
RT "Functional characterization of MpaG', the O-methyltransferase involved in
RT the biosynthesis of mycophenolic acid.";
RL ChemBioChem 16:565-569(2015).
RN [2]
RP FUNCTION, SUBCELLULAR LOCATION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=31209052; DOI=10.1073/pnas.1821932116;
RA Zhang W., Du L., Qu Z., Zhang X., Li F., Li Z., Qi F., Wang X., Jiang Y.,
RA Men P., Sun J., Cao S., Geng C., Qi F., Wan X., Liu C., Li S.;
RT "Compartmentalized biosynthesis of mycophenolic acid.";
RL Proc. Natl. Acad. Sci. U.S.A. 116:13305-13310(2019).
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:31209052). 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:31209052). 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:31209052).
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:31209052) (Probable).
CC {ECO:0000269|PubMed:31209052, ECO:0000305|PubMed:31209052}.
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:0000269|PubMed:31209052};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66669;
CC Evidence={ECO:0000269|PubMed:31209052};
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:0000269|PubMed:31209052};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66673;
CC Evidence={ECO:0000269|PubMed:31209052};
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:31209052}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000269|PubMed:31209052}; Single-pass membrane protein
CC {ECO:0000255}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; KM595305; AJG44382.1; -; Genomic_DNA.
DR AlphaFoldDB; A0A0B5KYT4; -.
DR SMR; A0A0B5KYT4; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0005789; C:endoplasmic reticulum membrane; IDA: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; IDA: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; IDA: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 SUPFAM; SSF48264; SSF48264; 1.
DR SUPFAM; SSF56281; SSF56281; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Endoplasmic reticulum; Heme; Iron; Membrane; Metal-binding; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..853
FT /note="Cytochrome P450 monooxygenase mpaDE'"
FT /evidence="ECO:0000255"
FT /id="PRO_0000451892"
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..853
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT BINDING 449
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 853 AA; 96352 MW; C5C758984E11E6EF CRC64;
MESLSLTWIT AIAVVLYLVQ RYVRSYWRLK DIPGPVLAKL TDLQRVWWVK TGRAHEFHRD
MHAMYGPIVR FGPNMVSVSD PRVIPTIYPS RPGFPKGDFY RTQKPYTRNK GAMPAVFNTQ
DEDLHKQLRS PIASLYSMTN VVRLEPLVDE TLTVLSKQLD ERFVGTNDKP FDLGDWLQYF
AFDSMGTLTF SRRYGFLEQG RDMHGILQEI WNFMTRVAVM GQIPWFDEIW NKNSFITLFK
RPTGFGVLKV VDNFISQRVS SRENDEKADE KDMLSQFLNI QASNPHSIMP WAPRAWTFSN
VMAGSDSTAN VMRTMMYNLL VDRDTLKSLR AELLEAESSN GLSRSLPSWD GVRSLPYLDA
CVLEALRLHP PFCLPFERVV PEGGITVCET YLPAGTVVGI SPYLANRDKQ TFGDDADKWR
PSRWLDLSRE DRVKLENSIL TFGAGRRTCL GKNIAILEIK KLFPMLLLNY EIEIVNPENY
QTTNAWFFRQ WGLHAVIRKL PAPERDDTIE QKASIPPALN IPPSSSTVDV RIIDSGTLLD
LRPDLFWTPD LPGLLKVTAP TYCFLISNGS RHVLFDLAVR QDWENLPPSI VAMIKSQTVI
QEPRNISDVL DSDESSLGIR SKDIEAIIWS HAHFDHIGDP STFPPSTELV VGPGIRDTHW
PGFPTNPDAI NLNTDIQGRN VREISFEKTQ KGATKIGSFD AMDYFGDGSF YLLDAAGHSV
GHIGALARVT TSPDSFVFMG GDSCHHAGVL RPTKYLPCPL DSGDTSLPCK SDSVFTLSPA
LPTDYTAALR TVENIKELDA CEDVFVVLAH DATLKGKVDF YPSKINDWKA KEYGKKTKWL
FYKDIENAIE GQK
