LOVD_ASPTE
ID LOVD_ASPTE Reviewed; 413 AA.
AC Q9Y7D1;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1999, sequence version 1.
DT 03-AUG-2022, entry version 75.
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 Name=lovD {ECO:0000303|PubMed:10334994};
OS Aspergillus terreus.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=33178;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, PATHWAY, DISRUPTION PHENOTYPE,
RP AND SUBUNIT.
RC STRAIN=ATCC 20542 / MF4845;
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 [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.
RC STRAIN=ATCC 20542 / MF4845; TISSUE=Mycelium {ECO:0000269|PubMed:10381407};
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.
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 [5]
RP FUNCTION, MUTAGENESIS OF SER-76, 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 [6]
RP FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY, BIOPHYSICOCHEMICAL PROPERTIES,
RP AND MUTAGENESIS OF CYS-40 AND CYS-60.
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 [7]
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 [8]
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 [9]
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 [10]
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 [11]
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 [12]
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 [13]
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 [14]
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).
RN [15]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEXES WITH MONACOLIN J;
RP LOVASTATIN AND SIMVASTATIN, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE,
RP MUTAGENESIS OF ASP-12; LYS-26; ALA-86; HIS-161; ALA-190; GLY-275 AND
RP VAL-334, AND BIOPHYSICOCHEMICAL 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 [16]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS), FUNCTION, PATHWAY, AND MUTAGENESIS
RP OF ILE-4; ALA-9; LYS-26; ARG-28; ILE-35; ASN-43; ASP-96; SER-109; ALA-123;
RP MET-157; SER-164; SER-172; LEU-174; ALA-178; ASN-191; LEU-192; GLN-241;
RP ALA-247; ARG-250; SER-256; ALA-261; GLY-275; GLN-297; TRP-355; LEU-361;
RP VAL-370; ALA-383; ASN-391 AND HIS-404.
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).
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; AF141925; AAD34555.1; -; Genomic_DNA.
DR PDB; 3HL9; X-ray; 3.40 A; A/B/C/D=1-413.
DR PDB; 3HLB; X-ray; 2.50 A; A/B/C/D=1-413.
DR PDB; 3HLC; X-ray; 2.00 A; A=1-413.
DR PDB; 3HLD; X-ray; 2.00 A; A=1-413.
DR PDB; 3HLE; X-ray; 2.06 A; A=1-413.
DR PDB; 3HLF; X-ray; 2.00 A; A=1-413.
DR PDB; 3HLG; X-ray; 2.01 A; A=1-413.
DR PDB; 4LCL; X-ray; 1.80 A; A/B=1-413.
DR PDB; 4LCM; X-ray; 3.19 A; A/B/C/D=1-413.
DR PDBsum; 3HL9; -.
DR PDBsum; 3HLB; -.
DR PDBsum; 3HLC; -.
DR PDBsum; 3HLD; -.
DR PDBsum; 3HLE; -.
DR PDBsum; 3HLF; -.
DR PDBsum; 3HLG; -.
DR PDBsum; 4LCL; -.
DR PDBsum; 4LCM; -.
DR AlphaFoldDB; Q9Y7D1; -.
DR SMR; Q9Y7D1; -.
DR PRIDE; Q9Y7D1; -.
DR KEGG; ag:AAD34555; -.
DR VEuPathDB; FungiDB:ATEG_09964; -.
DR BioCyc; MetaCyc:MON-18785; -.
DR BRENDA; 2.3.1.238; 536.
DR UniPathway; UPA00875; -.
DR EvolutionaryTrace; Q9Y7D1; -.
DR GO; GO:0016746; F:acyltransferase activity; IDA:UniProtKB.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IDA:UniProtKB.
DR GO; GO:0050832; P:defense response to fungus; IDA:UniProtKB.
DR GO; GO:0030639; P:polyketide biosynthetic process; IDA:UniProtKB.
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 3D-structure; Acyltransferase; Hydrolase; Transferase.
FT CHAIN 1..413
FT /note="Monacolin J acid methylbutanoyltransferase"
FT /id="PRO_0000430100"
FT ACT_SITE 76
FT /note="Acyl-ester intermediate"
FT /evidence="ECO:0000269|PubMed:17113998,
FT ECO:0000269|PubMed:19875080"
FT BINDING 73
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT BINDING 173
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT BINDING 188
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT BINDING 258
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT BINDING 366
FT /ligand="2-methylbutanoate"
FT /ligand_id="ChEBI:CHEBI:48946"
FT BINDING 388
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT BINDING 390
FT /ligand="monacolin J"
FT /ligand_id="ChEBI:CHEBI:79034"
FT MUTAGEN 4
FT /note="I->N: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with V-9; E-26; S-28; L-35; R-43; R-96; C-109;
FT P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 9
FT /note="A->V: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; E-26; S-28; L-35; R-43; R-96; C-109;
FT P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 12
FT /note="D->G: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with E-26; A-40; N-60; V-86; Y-161; T-190; S-275 and F-
FT 334."
FT /evidence="ECO:0000269|PubMed:19875080"
FT MUTAGEN 26
FT /note="K->E: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; S-28; L-35; R-43; R-96; C-109; P-
FT 123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:19875080,
FT ECO:0000269|PubMed:24727900"
FT MUTAGEN 26
FT /note="K->E: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; A-40; N-60; V-86; Y-161; T-190; S-275 and F-
FT 334."
FT /evidence="ECO:0000269|PubMed:19875080,
FT ECO:0000269|PubMed:24727900"
FT MUTAGEN 28
FT /note="R->S: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; L-35; R-43; R-96; C-109; P-
FT 123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 35
FT /note="I->L: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; R-43; R-96; C-109; P-
FT 123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 40
FT /note="C->A: Improves protein solubility upon
FT overexpression in E.coli. Strongly improves protein
FT solubility; when associated with N-60. Strongly increases
FT simvastatin synthase activity upon overexpression in
FT E.coli; when associated with G-12; E-26; N-60; V-86; Y-161;
FT T-190; S-275 and F-334."
FT /evidence="ECO:0000269|PubMed:18988191"
FT MUTAGEN 43
FT /note="N->R: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-96; C-109; P-
FT 123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 60
FT /note="C->A,N: Minor effect on protein solubility upon
FT overexpression in E.coli. Strongly improves protein
FT solubility; when associated with A-40. Strongly increases
FT simvastatin synthase activity upon overexpression in
FT E.coli; when associated with G-12; E-26; A-40; V-86; Y-161;
FT T-190; S-275 and F-334."
FT /evidence="ECO:0000269|PubMed:18988191"
FT MUTAGEN 76
FT /note="S->A,N: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:17113998"
FT MUTAGEN 86
FT /note="A->V: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; E-26; A-40; N-60; Y-161; T-190; S-275 and F-
FT 334."
FT /evidence="ECO:0000269|PubMed:19875080"
FT MUTAGEN 96
FT /note="D->R: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; C-109; P-
FT 123; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 109
FT /note="S->C: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96;."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 123
FT /note="A->P: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; V-157; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 157
FT /note="M->V: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; G-164; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 161
FT /note="H->Y: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; E-26; A-40; N-60; V-86; T-190; S-275 and F-334."
FT /evidence="ECO:0000269|PubMed:19875080"
FT MUTAGEN 164
FT /note="S->G: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; N-172; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 172
FT /note="S->N: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; F-174; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; V-383; M-
FT 355; L-361; I-370; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 174
FT /note="L->F: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; L-178; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 178
FT /note="A->L: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; G-191; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 190
FT /note="A->T: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; E-26; A-40; N-60; V-86; Y-161; S-275 and F-334."
FT /evidence="ECO:0000269|PubMed:19875080"
FT MUTAGEN 191
FT /note="N->G: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; I-192; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 192
FT /note="L->I: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; M-
FT 241; S-247; K-250; T-256; H-261; S-275; G-297; V-383; M-
FT 355; L-361; I-370; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 241
FT /note="Q->M: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; S-247; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 247
FT /note="A->S: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; K-250; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 250
FT /note="R->K: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; T-256; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 256
FT /note="S->T: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; H-261; S-275; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 261
FT /note="A->H: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; S-275; G-297; V-383; M-
FT 355; L-361; I-370; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 275
FT /note="G->S: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; G-297; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:19875080,
FT ECO:0000269|PubMed:24727900"
FT MUTAGEN 275
FT /note="G->S: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; E-26; A-40; N-60; V-86; Y-161; T-190 and F-334."
FT /evidence="ECO:0000269|PubMed:19875080,
FT ECO:0000269|PubMed:24727900"
FT MUTAGEN 297
FT /note="Q->G: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; M-355; L-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 334
FT /note="V->F: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli; when associated
FT with G-12; E-26; A-40; N-60; V-86; Y-161; T-190 and S-275."
FT /evidence="ECO:0000269|PubMed:19875080"
FT MUTAGEN 355
FT /note="W->M: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; G-297; M-
FT 361; I-370; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 361
FT /note="L->M: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; V-383; G-
FT 297; M-355; I-370; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 370
FT /note="V->I: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; G-297; M-
FT 355; M-361; V-383; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 383
FT /note="A->V: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; G-297; M-
FT 355; L-361; I-370; S-391 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 391
FT /note="N->S: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; G-297; M-
FT 355; M-361; I-370; V-383 and K-404."
FT /evidence="ECO:0000269|PubMed:24727900"
FT MUTAGEN 404
FT /note="H->K: Strongly increases simvastatin synthase
FT activity upon overexpression in E.coli and abolishes
FT activity with LovD-bound alpha-methylbutanoate; when
FT associated with N-4; V-9; E-26; S-28; L-35; R-43; R-96; C-
FT 109; P-123; V-157; G-164; N-172; F-174; L-178; G-191; I-
FT 192; M-241; S-247; K-250; T-256; H-261; S-275; V-383; G-
FT 297; M-355; M-361; I-370; V-383 and S-391."
FT /evidence="ECO:0000269|PubMed:24727900"
FT HELIX 4..10
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 13..26
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 31..39
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 45..53
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 75..77
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 78..91
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 101..103
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 106..109
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 113..117
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 123..126
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 134..138
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 147..149
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 151..159
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 160..164
FT /evidence="ECO:0007829|PDB:3HLD"
FT STRAND 165..167
FT /evidence="ECO:0007829|PDB:3HLD"
FT STRAND 173..176
FT /evidence="ECO:0007829|PDB:3HLD"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 182..184
FT /evidence="ECO:0007829|PDB:3HLG"
FT HELIX 191..203
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 207..214
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 217..219
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 223..226
FT /evidence="ECO:0007829|PDB:3HLC"
FT HELIX 228..230
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 232..237
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 241..244
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 246..248
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 251..254
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 257..259
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 268..270
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 276..288
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 291..294
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 296..302
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 309..320
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 323..325
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 327..330
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 337..339
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 341..348
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 352..354
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 361..365
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 366..368
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 369..374
FT /evidence="ECO:0007829|PDB:4LCL"
FT TURN 375..378
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 379..384
FT /evidence="ECO:0007829|PDB:4LCL"
FT STRAND 387..391
FT /evidence="ECO:0007829|PDB:4LCL"
FT HELIX 393..412
FT /evidence="ECO:0007829|PDB:4LCL"
SQ SEQUENCE 413 AA; 46037 MW; 731A140B6E609A24 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
LMFQPALEPR LEEQMNQHMD ASPHINYGGP MPMVLRRSFG LGGIIALEDL DGENWRRKGS
LTFGGGPNIV WQIDPKAGLC TLAFFQLEPW NDPVCRDLTR TFEHAIYAQY QQG
