3HLE

Simvastatin Synthase (LovD), from Aspergillus terreus, S5 mutant, S76A mutant, complex with monacolin J acid

Summary for 3HLE

• Entry DOI: 10.2210/pdb3hle/pdb
• Related: 1HLD
• Descriptor: Transesterase, (3R,5R)-3,5-dihydroxy-7-[(1S,2S,6R,8S,8aR)-8-hydroxy-2,6-dimethyl-1,2,6,7,8,8a-hexahydronaphthalen-1-yl]heptanoic acid, 2,3-DIHYDROXY-1,4-DITHIOBUTANE, ... (4 entities in total)
• Functional Keywords: alpha/beta hydrolase fold, transferase
• Biological source: Aspergillus terreus
• Total number of polymer chains: 1
• Total formula weight: 48640.35

Authors

Sawaya, M.R.,Yeates, T.O.,Pashkov, I.,Gao, X.,Tang, Y. (deposition date: 2009-05-27, release date: 2009-10-27, Last modification date: 2023-09-06)

Primary citation

Gao, X.,Xie, X.,Pashkov, I.,Sawaya, M.R.,Laidman, J.,Zhang, W.,Cacho, R.,Yeates, T.O.,Tang, Y.
Directed evolution and structural characterization of a simvastatin synthase
Chem.Biol., 16:1064-1074, 2009

PubMed Abstract: 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.

PubMed: 19875080
DOI: 10.1016/j.chembiol.2009.09.017

Experimental method

X-RAY DIFFRACTION (2.06 Å)