Enhancing the thermostability of glucose isomerase by protein engineering

Biotechnology (N Y). 1991 Aug;9(8):738-42. doi: 10.1038/nbt0891-738.

Abstract

We have engineered recombinant glucose isomerase (GI) from Actinoplanes missouriensis by site-directed mutagenesis to enhance its thermal stability in both the soluble and immobilized forms. Substitution of arginine for lysine at position 253, which lies at the dimer/dimer interface of the GI tetramer, produced the largest stabilization under model industrial conditions. We discuss our results in terms of a model in which chemical glycation of lysines by sugars in the industrial corn syrup substrate represents a major pathway of destabilization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldose-Ketose Isomerases*
  • Carbohydrate Epimerases / chemistry*
  • Carbohydrate Epimerases / genetics
  • Cloning, Molecular
  • Enzymes, Immobilized
  • Escherichia coli / genetics
  • Gram-Positive Bacteria / enzymology
  • Gram-Positive Bacteria / genetics
  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Mutagenesis
  • Protein Engineering
  • Solutions
  • Thermodynamics

Substances

  • Enzymes, Immobilized
  • Solutions
  • Carbohydrate Epimerases
  • Aldose-Ketose Isomerases
  • xylose isomerase