4LCM

Simvastatin Synthase (LOVD), from Aspergillus Terreus, LovD9 mutant (simh9014)

Summary for 4LCM

• Entry DOI: 10.2210/pdb4lcm/pdb
• Related: 3HL9 3HLB 3HLC 3HLD 3HLE 3HLF 3HLG 4LCL
• Descriptor: Transesterase (2 entities in total)
• Functional Keywords: laboratory-directed evolution, transesterase, transferase
• Biological source: Aspergillus terreus
• Total number of polymer chains: 4
• Total formula weight: 189084.03

Authors

Gao, X.,Sawaya, M.R.,Yeates, T.O.,Tang, Y. (deposition date: 2013-06-21, release date: 2014-04-02, Last modification date: 2023-09-20)

Primary citation

Jimenez-Oses, G.,Osuna, S.,Gao, X.,Sawaya, M.R.,Gilson, L.,Collier, S.J.,Huisman, G.W.,Yeates, T.O.,Tang, Y.,Houk, K.N.
The role of distant mutations and allosteric regulation on LovD active site dynamics.
Nat.Chem.Biol., 10:431-436, 2014

PubMed Abstract: Natural enzymes have evolved to perform their cellular functions under complex selective pressures, which often require their catalytic activities to be regulated by other proteins. We contrasted a natural enzyme, LovD, which acts on a protein-bound (LovF) acyl substrate, with a laboratory-generated variant that was transformed by directed evolution to accept instead a small free acyl thioester and no longer requires the acyl carrier protein. The resulting 29-mutant variant is 1,000-fold more efficient in the synthesis of the drug simvastatin than the wild-type LovD. This is to our knowledge the first nonpatent report of the enzyme currently used for the manufacture of simvastatin as well as the intermediate evolved variants. Crystal structures and microsecond-scale molecular dynamics simulations revealed the mechanism by which the laboratory-generated mutations free LovD from dependence on protein-protein interactions. Mutations markedly altered conformational dynamics of the catalytic residues, obviating the need for allosteric modulation by the acyl carrier LovF.

PubMed: 24727900
DOI: 10.1038/nchembio.1503

Experimental method

X-RAY DIFFRACTION (3.19 Å)