MPAH2_PENBR
ID MPAH2_PENBR Reviewed; 433 AA.
AC A0A0B5LB55;
DT 10-FEB-2021, integrated into UniProtKB/Swiss-Prot.
DT 10-FEB-2021, sequence version 2.
DT 03-AUG-2022, entry version 11.
DE RecName: Full=Type I acyl-CoA thioesterase mpaH' {ECO:0000303|PubMed:31209052};
DE EC=3.1.1.- {ECO:0000269|PubMed:31209052};
DE AltName: Full=Mycophenolic acid biosynthesis cluster protein H' {ECO:0000303|PubMed:25630520};
GN Name=mpaH' {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, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP SER-139, ACTIVE SITE, DISRUPTION PHENOTYPE, SUBCELLULAR LOCATION, AND
RP 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).
RN [3] {ECO:0007744|PDB:7DBI, ECO:0007744|PDB:7DBL}
RP X-RAY CRYSTALLOGRAPHY (1.84 ANGSTROMS) IN COMPLEX WITH MYCOPHENOLIC ACID,
RP FUNCTION, ACTIVE SITE, CATALYTIC ACTIVITY, MUTAGENESIS OF SER-139; ASP-163
RP AND HIS-365, SUBUNIT, AND PATHWAY.
RX PubMed=33843134; DOI=10.1111/febs.15874;
RA You C., Li F., Zhang X., Ma L., Zhang Y.Z., Zhang W., Li S.;
RT "Structural basis for substrate specificity of the peroxisomal acyl-CoA
RT hydrolase MpaH' involved in mycophenolic acid biosynthesis.";
RL FEBS J. 288:5768-5780(2021).
CC -!- FUNCTION: Type I acyl-CoA thioesterase; 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,
CC PubMed:33843134). MpaH' acts as a peroxisomal acyl-CoA hydrolase that
CC converts MPA-CoA into the final product MPA (PubMed:31209052,
CC PubMed:33843134). The first step of the pathway is the synthesis of 5-
CC methylorsellinic acid (5MOA) by the cytosolic polyketide synthase mpaC.
CC 5MOA is then converted to the phthalide compound 5,7-dihydroxy-4,6-
CC dimethylphthalide (DHMP) by the endoplasmic reticulum-bound cytochrome
CC P450 monooxygenase mpaDE. MpaDE first catalyzes hydroxylation of 5-MOA
CC to 4,6-dihydroxy-2-(hydroxymethyl)-3-methylbenzoic acid (DHMB). MpaDE
CC then acts as a lactone synthase that catalyzes the ring closure to
CC convert DHMB into DHMP. The next step is the prenylation of DHMP by the
CC Golgi apparatus-associated prenyltransferase mpaA to yield farnesyl-
CC DHMP (FDHMP). The ER-bound oxygenase mpaB then mediates the oxidative
CC cleavage the C19-C20 double bond in FDHMP to yield FDHMP-3C via a
CC mycophenolic aldehyde intermediate. The O-methyltransferase mpaG
CC catalyzes the methylation of FDHMP-3C to yield MFDHMP-3C. After the
CC cytosolic methylation of FDHMP-3C, MFDHMP-3C enters into peroxisomes
CC probably via free diffusion due to its low molecular weight. Upon a
CC peroxisomal CoA ligation reaction, catalyzed by a beta-oxidation
CC component enzyme acyl-CoA ligase ACL891, MFDHMP-3C-CoA would then be
CC restricted to peroxisomes for the following beta-oxidation pathway
CC steps. The peroxisomal beta-oxidation machinery than converts MFDHMP-
CC 3C-CoA into MPA_CoA, via a beta-oxidation chain-shortening process.
CC Finally mpaH acts as a peroxisomal acyl-CoA hydrolase with high
CC substrate specificity toward MPA-CoA to release the final product MPA
CC (PubMed:31209052) (Probable). {ECO:0000269|PubMed:31209052,
CC ECO:0000269|PubMed:33843134, ECO:0000305|PubMed:31209052}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + mycophenolyl-CoA = CoA + H(+) + mycophenolate;
CC Xref=Rhea:RHEA:66704, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57287, ChEBI:CHEBI:62932, ChEBI:CHEBI:167447;
CC Evidence={ECO:0000269|PubMed:31209052, ECO:0000269|PubMed:33843134};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66705;
CC Evidence={ECO:0000269|PubMed:31209052, ECO:0000269|PubMed:33843134};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=722.3 uM for acetyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=351.5 uM for propionyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=76.37 uM for malonyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=201.9 uM for isobutyryl-CoA {ECO:0000269|PubMed:31209052};
CC KM=780.83 uM for isovaleryl-CoA {ECO:0000269|PubMed:31209052};
CC KM=217.9 uM for benzoyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=143.0 uM for N-decanoyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=260.2 uM for lauroyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=577.0 uM for palmitoyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=316.6 uM for arachidonoyl-CoA {ECO:0000269|PubMed:31209052};
CC KM=382.6 uM for DMMPA-CoA {ECO:0000269|PubMed:31209052};
CC KM=117.5 uM for MPA-CoA {ECO:0000269|PubMed:31209052};
CC Note=The kcat values are 54.98 min(-1) for acetyl-CoA, 24.58 min(-1)
CC for propionyl-CoA, 12.94 min(-1) for malonyl-CoA, 7.688 min(-1) for
CC isobutyryl-CoA, 5.718 min(-1) for isovaleryl-CoA, 3.891 min(-1) for
CC benzoyl-CoA, 136.5 min(-1) for N-decanoyl-CoA, 99.11 min(-1) for
CC lauroyl-CoA, 23.19 min(-1) for palmitoyl-CoA, 47.35 min(-1) for
CC arachidonoyl-CoA, 4438 min(-1) for DMMPA-CoA and 9578 min(-1) for
CC MPA-CoA. {ECO:0000269|PubMed:31209052};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:31209052, ECO:0000269|PubMed:33843134}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:33843134}.
CC -!- SUBCELLULAR LOCATION: Peroxisome matrix {ECO:0000269|PubMed:31209052}.
CC Note=The mpaH' location in peroxisomes is required for the unique
CC cooperation between biosynthetic and beta-oxidation catabolism
CC machineries to produce final MPA. {ECO:0000269|PubMed:31209052}.
CC -!- DISRUPTION PHENOTYPE: Retains the ability to produce MPA, albeit with
CC an about 50% lower yield, and accumulates 2 compounds (MFDHMP-d4 and
CC MFDHMP-d5) with an even shorter isoprenyl chain than MPA.
CC {ECO:0000269|PubMed:31209052}.
CC -!- SIMILARITY: Belongs to the AB hydrolase superfamily. MpaH hydrolase
CC family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AJG44385.1; Type=Erroneous gene model prediction; Evidence={ECO:0000305};
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DR EMBL; KM595305; AJG44385.1; ALT_SEQ; Genomic_DNA.
DR PDB; 7DBI; X-ray; 1.99 A; A/B/C/D=1-433.
DR PDB; 7DBL; X-ray; 1.84 A; A/B/C/D=1-433.
DR PDBsum; 7DBI; -.
DR PDBsum; 7DBL; -.
DR AlphaFoldDB; A0A0B5LB55; -.
DR SMR; A0A0B5LB55; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0005782; C:peroxisomal matrix; IDA:GO_Central.
DR GO; GO:0016787; F:hydrolase activity; IDA:GO_Central.
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; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR000073; AB_hydrolase_1.
DR Pfam; PF12697; Abhydrolase_6; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Peroxisome.
FT CHAIN 1..433
FT /note="Type I acyl-CoA thioesterase mpaH'"
FT /id="PRO_0000451895"
FT REGION 58..246
FT /note="Abhydrolase domain"
FT /evidence="ECO:0000255"
FT ACT_SITE 139
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:31209052,
FT ECO:0000269|PubMed:33843134"
FT ACT_SITE 163
FT /evidence="ECO:0000269|PubMed:33843134,
FT ECO:0000305|PubMed:31209052"
FT ACT_SITE 365
FT /evidence="ECO:0000269|PubMed:33843134,
FT ECO:0000305|PubMed:31209052"
FT BINDING 60
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:33843134,
FT ECO:0007744|PDB:7DBL"
FT BINDING 140
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:33843134,
FT ECO:0007744|PDB:7DBL"
FT MUTAGEN 139
FT /note="S->A: Abolishes the acyl-CoA thioesterase activity."
FT /evidence="ECO:0000269|PubMed:31209052,
FT ECO:0000269|PubMed:33843134"
FT MUTAGEN 163
FT /note="D->A: Loses 49.5% of the acyl-CoA thioesterase
FT activity."
FT /evidence="ECO:0000269|PubMed:33843134"
FT MUTAGEN 365
FT /note="H->A: Abolishes the acyl-CoA thioesterase activity."
FT /evidence="ECO:0000269|PubMed:33843134"
FT STRAND 4..14
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 22..24
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 25..27
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 33..41
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 52..57
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 64..67
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 68..76
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 83..89
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 95..100
FT /evidence="ECO:0007829|PDB:7DBL"
FT TURN 101..104
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 111..124
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 126..128
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 133..138
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 141..151
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 155..163
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 182..188
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 192..195
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 196..202
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 204..207
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 212..221
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 222..226
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 229..231
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 234..241
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 252..256
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 258..265
FT /evidence="ECO:0007829|PDB:7DBL"
FT TURN 275..277
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 284..287
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 292..294
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 297..300
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 302..309
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 310..313
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 316..323
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 330..339
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 350..352
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 355..360
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 367..370
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 372..401
FT /evidence="ECO:0007829|PDB:7DBL"
FT STRAND 406..408
FT /evidence="ECO:0007829|PDB:7DBL"
FT HELIX 412..416
FT /evidence="ECO:0007829|PDB:7DBL"
SQ SEQUENCE 433 AA; 49147 MW; 6FA2BB01D079BE58 CRC64;
MSTEKFTITE HLVPGSHIRE YPGSTVNQED VLKIHVKQYT PKREGPVPDD AITFIATHGV
GLPKELYEPL WDELLDQASG FHIRAIWMAD VASMNQSGIH NEDKLSMDCS WMDHARDLLL
MINHFRDQMP RPLVGIGHSF GGNIITNLAY LHPRLFTTLL LLDPLIQLSP PSLGFGTDAP
SAINYTLWRD DVWPSREVAI RANRAIMQGM DPRCLDRMTK HFFRDLPTPL YPDVEAIKAL
FGTTADSTTT PVTLTTPKYH ELVAQIRQNF NARDPKTGRI EVPRDTHADM DPLVAYIPLY
RPEPRSTFRR LETLRPSCLW VIAGATFLNI DEIREGVKIC GSGIGGSGGV PDGRVREVVL
PGFGHLMPFQ EVKTVAETCI VWLQQEMDRF RQTERQWKED RDGKSHLAVE ENWYKVLKPI
PSGRKKRNDK GKL
