Immobilization of UDP-galactose 4-epimerase from Escherichia coli on the yeast cell surface

Biosci Biotechnol Biochem. 2006 Sep;70(9):2303-6. doi: 10.1271/bbb.60134. Epub 2006 Sep 7.

Abstract

UDP-galactose 4-epimerase (EC 5.1.3.2, Gal E) from Escherichia coli catalyzes the reversible reaction between UDP-galactose and UDP-glucose. In this study, the Gal E gene from E. coli, coding UDP-galactose 4-epimerase, was cloned into pYD1 plasmid and then transformed into Saccharomyces cerevisiae EBY100 for expression of Gal E on the cell surface. Enzyme activity analyses with EBY100 cells showed that the enzyme displayed on the yeast cell surface was very active in the conversion between UDP-Glc and UDP-Gal. It took about 3 min to reach equilibrium from UDP-galactose to UDP-glucose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Electrophoresis, Capillary
  • Enzymes, Immobilized / genetics
  • Enzymes, Immobilized / metabolism*
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Polymerase Chain Reaction
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • UDPglucose 4-Epimerase / genetics
  • UDPglucose 4-Epimerase / metabolism*
  • Uridine Diphosphate Galactose / metabolism
  • Uridine Diphosphate Glucose / metabolism

Substances

  • DNA, Bacterial
  • Enzymes, Immobilized
  • Uridine Diphosphate Galactose
  • UDPglucose 4-Epimerase
  • Uridine Diphosphate Glucose