Probing the substrate specificity for lipases. II. Kinetic and modeling studies on the molecular recognition of 2-arylpropionic esters by Candida rugosa and Rhizomucor miehei lipases

Biochim Biophys Acta. 1997 Feb 8;1337(2):302-10. doi: 10.1016/s0167-4838(96)00181-1.

Abstract

Racemic arylpropionic esters 1-3, precursors of therapeutically important non-steroidal antiinflammatory drugs, were subjected to hydrolyses in the presence of either Candida rugosa or Rhizomucor miehei crude lipases. The hydrolyses of 1 and 2 proved to be highly enantioselective, whereas 3 was not transformed at all. Both the substrate specificity and the enantioselectivity of these lipases were explained through a molecular modeling study involving docking experiments between 1-3 and the amino acids forming the enzymes active-sites, whose three dimensional structures were obtained from X-ray crystallographic data, followed by extensive conformational analysis on their computer-generated complexes. The results of this study also account for the high enantioselective and good yielding hydrolysis of 3 (as the corresponding 2-chloroethyl ester) catalyzed by CRL pretreated with 2-propanol, recently reported in the literature, and lead to admit that such a treatment may operate very deep conformational changes on the amino acids of the enzyme active-site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Candida / enzymology*
  • Crystallography, X-Ray
  • Kinetics
  • Lipase / chemistry
  • Lipase / metabolism*
  • Models, Molecular
  • Molecular Structure
  • Mucorales / enzymology*
  • Propionates / chemistry
  • Propionates / metabolism*
  • Protein Conformation
  • Substrate Specificity

Substances

  • Propionates
  • Lipase