Effect of mutations in Candida antarctica B lipase

Chem Phys Lipids. 1998 Jun;93(1-2):95-101. doi: 10.1016/s0009-3084(98)00032-2.

Abstract

Three variants of the Candida antarctica B lipase have been constructed and characterized. The variant containing the T103G mutation, which introduces the consensus sequence G-X-S-X-G found in most other known lipases, shows an increased thermostability but retains only half the specific activity of the native enzyme. Also in ester synthesis the activity is lowered but the specificity and enantioselectivity remains unchanged. The W104H mutant, in which more space is introduced into the active site, has more dramatically changed properties. Both the thermostability and the specific activity are slightly reduced but the activity and specificity in ester synthesis is highly different from the native enzyme. In general, the activity is very low and the enantioselectivity is, furthermore, highly reduced. Finally, the mutation M72L was introduced to increase the oxidation stability of the enzyme. This variant did exhibit an increased resistance towards oxidation but the thermostability was, unfortunately, also reduced.

MeSH terms

  • Candida / enzymology*
  • Enzyme Stability
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / genetics*
  • Enzymes, Immobilized / metabolism*
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism*
  • Hot Temperature
  • Lipase / chemistry
  • Lipase / genetics*
  • Lipase / metabolism*
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Mutation*
  • Oxidation-Reduction
  • Stereoisomerism
  • Substrate Specificity

Substances

  • Enzymes, Immobilized
  • Fungal Proteins
  • Lipase