Enantioselectivity of recombinant Rhizomucor miehei lipase in the ring opening of oxazolin-5(4H)-ones

Protein Eng. 2001 Apr;14(4):269-78. doi: 10.1093/protein/14.4.269.

Abstract

Enantioselectivity of enzyme catalysis is often rationalized via active site models. These models are constructed on the basis of comparing the enantiomeric excess of product observed in a series of reactions which are conducted with a range of homologous substrates, typically carrying various side chain substitutions. Surprisingly the practical application of these simple but informative 'pocket size' models has been rarely tested in genetic engineering experiments. In this paper we report the construction, purification and enantioselectivity of two recombinant Rhizomucor miehei lipases which were designed to check the validity of such a model in reactions of ring opening of oxazolin-5(4H)-ones.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalytic Domain
  • Cloning, Molecular
  • Computer Simulation
  • Fungal Proteins
  • Hydrogen Bonding
  • Kinetics
  • Lipase / genetics
  • Lipase / metabolism*
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Oxazoles / chemistry
  • Oxazoles / metabolism*
  • Oxazolone
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Rhizomucor / enzymology*
  • Stereoisomerism

Substances

  • Fungal Proteins
  • Oxazoles
  • Recombinant Proteins
  • Oxazolone
  • Lipase