3HLE

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

3HLE の概要

• エントリーDOI: 10.2210/pdb3hle/pdb
• 関連するPDBエントリー: 1HLD
• 分子名称: 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)
• 機能のキーワード: alpha/beta hydrolase fold, transferase
• 由来する生物種: Aspergillus terreus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 48640.35

構造登録者

Sawaya, M.R.,Yeates, T.O.,Pashkov, I.,Gao, X.,Tang, Y. (登録日: 2009-05-27, 公開日: 2009-10-27, 最終更新日: 2023-09-06)

主引用文献

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

実験手法

X-RAY DIFFRACTION (2.06 Å)