LOVD_ASPTN
ID LOVD_ASPTN Reviewed; 413 AA.
AC Q0C8M0;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 17-OCT-2006, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=Monacolin J acid methylbutanoyltransferase {ECO:0000303|PubMed:10334994};
DE EC=2.3.1.238 {ECO:0000269|PubMed:18988191, ECO:0000269|PubMed:19530726, ECO:0000269|PubMed:19875080};
DE AltName: Full=Lovastatin biosynthesis cluster protein D {ECO:0000303|PubMed:10334994};
DE AltName: Full=Lovastatin hydrolase;
DE AltName: Full=Simvastatin synthase LovD {ECO:0000303|PubMed:19875080};
DE Short=SV synthase {ECO:0000303|PubMed:19875080};
GN ORFNames=ATEG_09964;
OS Aspergillus terreus (strain NIH 2624 / FGSC A1156).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=341663;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=NIH 2624 / FGSC A1156;
RA Birren B.W., Lander E.S., Galagan J.E., Nusbaum C., Devon K., Henn M.,
RA Ma L.-J., Jaffe D.B., Butler J., Alvarez P., Gnerre S., Grabherr M.,
RA Kleber M., Mauceli E.W., Brockman W., Rounsley S., Young S.K., LaButti K.,
RA Pushparaj V., DeCaprio D., Crawford M., Koehrsen M., Engels R.,
RA Montgomery P., Pearson M., Howarth C., Larson L., Luoma S., White J.,
RA Alvarado L., Kodira C.D., Zeng Q., Oleary S., Yandava C., Denning D.W.,
RA Nierman W.C., Milne T., Madden K.;
RT "Annotation of the Aspergillus terreus NIH2624 genome.";
RL Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP BIOTECHNOLOGY.
RX PubMed=6933445; DOI=10.1073/pnas.77.7.3957;
RA Alberts A.W., Chen J., Kuron G., Hunt V., Huff J., Hoffman C., Rothrock J.,
RA Lopez M., Joshua H., Harris E., Patchett A., Monaghan R., Currie S.,
RA Stapley E., Albers-Schonberg G., Hensens O., Hirshfield J., Hoogsteen K.,
RA Liesch J., Springer J.;
RT "Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-
RT coenzyme A reductase and a cholesterol-lowering agent.";
RL Proc. Natl. Acad. Sci. U.S.A. 77:3957-3961(1980).
RN [3]
RP FUNCTION.
RX PubMed=10381407; DOI=10.1016/s1074-5521(99)80061-1;
RA Hendrickson L., Davis C.R., Roach C., Nguyen D.K., Aldrich T., McAda P.C.,
RA Reeves C.D.;
RT "Lovastatin biosynthesis in Aspergillus terreus: characterization of
RT blocked mutants, enzyme activities and a multifunctional polyketide
RT synthase gene.";
RL Chem. Biol. 6:429-439(1999).
RN [4]
RP FUNCTION, PATHWAY, DISRUPTION PHENOTYPE, AND SUBUNIT.
RX PubMed=10334994; DOI=10.1126/science.284.5418.1368;
RA Kennedy J., Auclair K., Kendrew S.G., Park C., Vederas J.C.,
RA Hutchinson C.R.;
RT "Modulation of polyketide synthase activity by accessory proteins during
RT lovastatin biosynthesis.";
RL Science 284:1368-1372(1999).
RN [5]
RP FUNCTION.
RX PubMed=12929390; DOI=10.1039/b207721c;
RA Sorensen J.L., Auclair K., Kennedy J., Hutchinson C.R., Vederas J.C.;
RT "Transformations of cyclic nonaketides by Aspergillus terreus mutants
RT blocked for lovastatin biosynthesis at the lovA and lovC genes.";
RL Org. Biomol. Chem. 1:50-59(2003).
RN [6]
RP FUNCTION, ACTIVE SITE, AND BIOTECHNOLOGY.
RX PubMed=17113998; DOI=10.1016/j.chembiol.2006.09.008;
RA Xie X., Watanabe K., Wojcicki W.A., Wang C.C., Tang Y.;
RT "Biosynthesis of lovastatin analogs with a broadly specific
RT acyltransferase.";
RL Chem. Biol. 13:1161-1169(2006).
RN [7]
RP BIOTECHNOLOGY, FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=19875080; DOI=10.1016/j.chembiol.2009.09.017;
RA Gao X., Xie X., Pashkov I., Sawaya M.R., Laidman J., Zhang W., Cacho R.,
RA Yeates T.O., Tang Y.;
RT "Directed evolution and structural characterization of a simvastatin
RT synthase.";
RL Chem. Biol. 16:1064-1074(2009).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=18988191; DOI=10.1002/bit.22028;
RA Xie X., Pashkov I., Gao X., Guerrero J.L., Yeates T.O., Tang Y.;
RT "Rational improvement of simvastatin synthase solubility in Escherichia
RT coli leads to higher whole-cell biocatalytic activity.";
RL Biotechnol. Bioeng. 102:20-28(2009).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, AND INTERACTION WITH LOVF.
RX PubMed=19530726; DOI=10.1021/ja903203g;
RA Xie X., Meehan M.J., Xu W., Dorrestein P.C., Tang Y.;
RT "Acyltransferase mediated polyketide release from a fungal megasynthase.";
RL J. Am. Chem. Soc. 131:8388-8389(2009).
RN [10]
RP FUNCTION.
RX PubMed=19900898; DOI=10.1126/science.1175602;
RA Ma S.M., Li J.W., Choi J.W., Zhou H., Lee K.K., Moorthie V.A., Xie X.,
RA Kealey J.T., Da Silva N.A., Vederas J.C., Tang Y.;
RT "Complete reconstitution of a highly reducing iterative polyketide
RT synthase.";
RL Science 326:589-592(2009).
RN [11]
RP FUNCTION.
RX PubMed=21069965; DOI=10.1021/bi1014776;
RA Meehan M.J., Xie X., Zhao X., Xu W., Tang Y., Dorrestein P.C.;
RT "FT-ICR-MS characterization of intermediates in the biosynthesis of the
RT alpha-methylbutyrate side chain of lovastatin by the 277 kDa polyketide
RT synthase LovF.";
RL Biochemistry 50:287-299(2011).
RN [12]
RP FUNCTION.
RX PubMed=21495633; DOI=10.1021/ja201138v;
RA Barriuso J., Nguyen D.T., Li J.W., Roberts J.N., MacNevin G., Chaytor J.L.,
RA Marcus S.L., Vederas J.C., Ro D.K.;
RT "Double oxidation of the cyclic nonaketide dihydromonacolin L to monacolin
RT J by a single cytochrome P450 monooxygenase, LovA.";
RL J. Am. Chem. Soc. 133:8078-8081(2011).
RN [13]
RP FUNCTION.
RX PubMed=22733743; DOI=10.1073/pnas.1113029109;
RA Ames B.D., Nguyen C., Bruegger J., Smith P., Xu W., Ma S., Wong E.,
RA Wong S., Xie X., Li J.W., Vederas J.C., Tang Y., Tsai S.C.;
RT "Crystal structure and biochemical studies of the trans-acting polyketide
RT enoyl reductase LovC from lovastatin biosynthesis.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:11144-11149(2012).
RN [14]
RP FUNCTION.
RX PubMed=23653178; DOI=10.1002/anie.201302406;
RA Xu W., Chooi Y.H., Choi J.W., Li S., Vederas J.C., Da Silva N.A., Tang Y.;
RT "LovG: the thioesterase required for dihydromonacolin L release and
RT lovastatin nonaketide synthase turnover in lovastatin biosynthesis.";
RL Angew. Chem. Int. Ed. Engl. 52:6472-6475(2013).
RN [15]
RP FUNCTION, AND PATHWAY.
RX PubMed=24727900; DOI=10.1038/nchembio.1503;
RA Jimenez-Oses G., Osuna S., Gao X., Sawaya M.R., Gilson L., Collier S.J.,
RA Huisman G.W., Yeates T.O., Tang Y., Houk K.N.;
RT "The role of distant mutations and allosteric regulation on LovD active
RT site dynamics.";
RL Nat. Chem. Biol. 10:431-436(2014).
RN [16]
RP BIOTECHNOLOGY.
RX PubMed=29236027; DOI=10.3390/ijms18122690;
RA Chen M.C., Tsai Y.C., Tseng J.H., Liou J.J., Horng S., Wen H.C., Fan Y.C.,
RA Zhong W.B., Hsu S.P.;
RT "Simvastatin inhibits cell proliferation and migration in human anaplastic
RT thyroid cancer.";
RL Int. J. Mol. Sci. 18:0-0(2017).
RN [17]
RP BIOTECHNOLOGY.
RX PubMed=29932104; DOI=10.3390/ijms19071834;
RA Zhong W.B., Tsai Y.C., Chin L.H., Tseng J.H., Tang L.W., Horng S.,
RA Fan Y.C., Hsu S.P.;
RT "A synergistic anti-cancer effect of troglitazone and lovastatin in a human
RT anaplastic thyroid cancer cell line and in a mouse xenograft model.";
RL Int. J. Mol. Sci. 19:0-0(2018).
CC -!- FUNCTION: Monacolin J acid methylbutanoyltransferase; part of the gene
CC cluster that mediates the biosynthesis of lovastatin (also known as
CC mevinolin, mevacor or monacolin K), a hypolipidemic inhibitor of (3S)-
CC hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase (HMGR)
CC (PubMed:10334994, PubMed:12929390, PubMed:21495633). The first step in
CC the biosynthesis of lovastatin is the production of dihydromonacolin L
CC acid by the lovastatin nonaketide synthase lovB and the trans-acting
CC enoyl reductase lovC via condensation of one acetyl-CoA unit and 8
CC malonyl-CoA units (PubMed:10334994, PubMed:10381407, PubMed:19900898,
CC PubMed:22733743). Dihydromonacolin L acid is released from lovB by the
CC thioesterase lovG (PubMed:23653178). Next, dihydromonacolin L acid is
CC oxidized by the dihydromonacolin L monooxygenase lovA twice to form
CC monacolin J acid (PubMed:12929390, PubMed:21495633). The 2-
CC methylbutyrate moiety of lovastatin is synthesized by the lovastatin
CC diketide synthase lovF via condensation of one acetyl-CoA unit and one
CC malonyl-CoA unit (PubMed:19530726, PubMed:21069965). Finally, the
CC covalent attachment of this moiety to monacolin J acid is catalyzed by
CC the transesterase lovD to yield lovastatin (PubMed:10334994,
CC PubMed:17113998, PubMed:18988191, PubMed:19875080, PubMed:24727900).
CC LovD has broad substrate specificity and can also convert monacolin J
CC to simvastatin using alpha-dimethylbutanoyl-S-methyl-3-
CC mercaptopropionate (DMB-S-MMP) as the thioester acyl donor, and can
CC also catalyze the reverse reaction and function as hydrolase in vitro
CC (PubMed:19875080). LovD has much higher activity with LovF-bound 2-
CC methylbutanoate than with free diketide substrates (PubMed:21069965).
CC {ECO:0000269|PubMed:10334994, ECO:0000269|PubMed:10381407,
CC ECO:0000269|PubMed:12929390, ECO:0000269|PubMed:17113998,
CC ECO:0000269|PubMed:18988191, ECO:0000269|PubMed:19530726,
CC ECO:0000269|PubMed:19875080, ECO:0000269|PubMed:19900898,
CC ECO:0000269|PubMed:21069965, ECO:0000269|PubMed:21495633,
CC ECO:0000269|PubMed:22733743, ECO:0000269|PubMed:23653178,
CC ECO:0000269|PubMed:24727900}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-2-methylbutanoyl-[2-methylbutanoate polyketide synthase] +
CC monacolin J carboxylate = holo-[2-methylbutanoate polyketide
CC synthase] + lovastatin carboxylate; Xref=Rhea:RHEA:43064, Rhea:RHEA-
CC COMP:10260, Rhea:RHEA-COMP:10261, ChEBI:CHEBI:64479,
CC ChEBI:CHEBI:79035, ChEBI:CHEBI:79038, ChEBI:CHEBI:82764;
CC EC=2.3.1.238; Evidence={ECO:0000269|PubMed:18988191,
CC ECO:0000269|PubMed:19530726, ECO:0000269|PubMed:19875080};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.78 mM for monacolin J {ECO:0000269|PubMed:18988191,
CC ECO:0000269|PubMed:19875080};
CC KM=0.67 mM for alpha-dimethylbutanoyl-S-methyl-3-mercaptopropionate
CC {ECO:0000269|PubMed:18988191, ECO:0000269|PubMed:19875080};
CC Note=kcat is 0.62 min(-1) for simvastatin synthesis.;
CC -!- PATHWAY: Polyketide biosynthesis; lovastatin biosynthesis.
CC {ECO:0000269|PubMed:10334994, ECO:0000269|PubMed:24727900}.
CC -!- SUBUNIT: Interacts with LovF. {ECO:0000269|PubMed:10334994,
CC ECO:0000269|PubMed:19530726}.
CC -!- DISRUPTION PHENOTYPE: Loss of lovastatin biosynthesis.
CC {ECO:0000269|PubMed:10334994}.
CC -!- BIOTECHNOLOGY: Lovastatin acts as a hypolipidemic agent that works as
CC inhibitor of (3S)-hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase
CC (HMGR) which reduces HMG-CoA to mevalonate and is the key step in
CC cholesterol biosynthesis (PubMed:6933445). Lovastatin, simvastatin and
CC related compounds are widely used to treat hypercholesteremia and
CC reduce the risk of cardiovascular disease (PubMed:6933445).
CC Furthermore, statins such as lovastatin were found to be anticancer
CC agents (PubMed:29236027, PubMed:29932104).
CC {ECO:0000269|PubMed:29236027, ECO:0000269|PubMed:29932104,
CC ECO:0000269|PubMed:6933445}.
CC -!- MISCELLANEOUS: Directed evolution toward higher catalytic activity with
CC free diketides led to an enzyme with 1000-fold higher activity in
CC simvastatin synthesis, due to numerous mutations that affect protein
CC folding and promote optimal alignment of the residues that are
CC important for substrate binding and catalysis.
CC {ECO:0000305|PubMed:24727900}.
CC -!- SIMILARITY: Belongs to the class-A beta-lactamase family.
CC {ECO:0000305}.
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DR EMBL; CH476609; EAU29413.1; -; Genomic_DNA.
DR RefSeq; XP_001209266.1; XM_001209266.1.
DR AlphaFoldDB; Q0C8M0; -.
DR SMR; Q0C8M0; -.
DR EnsemblFungi; EAU29413; EAU29413; ATEG_09964.
DR GeneID; 4319610; -.
DR VEuPathDB; FungiDB:ATEG_09964; -.
DR eggNOG; ENOG502S4UR; Eukaryota.
DR HOGENOM; CLU_020027_11_1_1; -.
DR OMA; WAGQMVE; -.
DR OrthoDB; 1268012at2759; -.
DR UniPathway; UPA00875; -.
DR Proteomes; UP000007963; Unassembled WGS sequence.
DR GO; GO:0016746; F:acyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.40.710.10; -; 1.
DR InterPro; IPR001466; Beta-lactam-related.
DR InterPro; IPR012338; Beta-lactam/transpept-like.
DR Pfam; PF00144; Beta-lactamase; 1.
DR SUPFAM; SSF56601; SSF56601; 1.
PE 1: Evidence at protein level;
KW Acyltransferase; Hydrolase; Reference proteome; Transferase.
FT CHAIN 1..413
FT /note="Monacolin J acid methylbutanoyltransferase"
FT /id="PRO_0000449660"
FT ACT_SITE 76
FT /note="Acyl-ester intermediate"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 73
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 173
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 188
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 258
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 366
FT /ligand="2-methylbutanoate"
FT /ligand_id="ChEBI:CHEBI:48946"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 388
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
FT BINDING 390
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT /evidence="ECO:0000250|UniProtKB:Q9Y7D1"
SQ SEQUENCE 413 AA; 46056 MW; 33DD09B7BF0D2732 CRC64;
MGSIIDAAAA ADPVVLMETA FRKAVKSRQI PGAVIMARDC SGNLNYTRCF GARTVRRDEC
NQLPPLQVDT PCRLASATKL LTTIMALQCM ERGLVDLDET VDRLLPDLSA MPVLEGFDDA
GNARLRERRG KITLRHLLTH TSGLSYVFLH PLLREYMAQG HLQSAEKFGI QSRLAPPAVN
DPGAEWIYGA NLDWAGKLVE RATGLDLEQY LQENICAPLG ITDMTFKLQQ RPDMLARRAD
QTHRNSADGR LRYDDSVYFR ADGEECFGGQ GVFSGPGSYM KVLHSLLKRD GLLLQPQTVD
MMFQPALEPR LEEQMNQHMD ASPHINYGGP MPMVLRRSFG LGGIIALEDL DGEDWRRKGS
LTFGGGPNIV WQIDPKAGLC TLAFFQLEPW NDPVCRDLTR TFEHAIYAQY QQG
