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PDBsum entry 3hlc
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References listed in PDB file
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Key reference
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Title
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Directed evolution and structural characterization of a simvastatin synthase.
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Authors
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X.Gao,
X.Xie,
I.Pashkov,
M.R.Sawaya,
J.Laidman,
W.Zhang,
R.Cacho,
T.O.Yeates,
Y.Tang.
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Ref.
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Chem Biol, 2009,
16,
1064-1074.
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PubMed id
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Abstract
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Enzymes from natural product biosynthetic pathways are attractive candidates for
creating tailored biocatalysts to produce semisynthetic pharmaceutical
compounds. LovD is an acyltransferase that converts the inactive monacolin J
acid (MJA) into the cholesterol-lowering lovastatin. LovD can also synthesize
the blockbuster drug simvastatin using MJA and a synthetic alpha-dimethylbutyryl
thioester, albeit with suboptimal properties as a biocatalyst. Here we used
directed evolution to improve the properties of LovD toward semisynthesis of
simvastatin. Mutants with improved catalytic efficiency, solubility, and thermal
stability were obtained, with the best mutant displaying an approximately
11-fold increase in an Escherichia coli-based biocatalytic platform. To
understand the structural basis of LovD enzymology, seven X-ray crystal
structures were determined, including the parent LovD, an improved mutant G5,
and G5 cocrystallized with ligands. Comparisons between the structures reveal
that beneficial mutations stabilize the structure of G5 in a more compact
conformation that is favorable for catalysis.
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