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Biotechnology Letters

, Volume 21, Issue 12, pp 1131–1135 | Cite as

Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli

  • Xi Chen
  • Przemyslaw Kowal
  • Sarah Hamad
  • Hongni Fan
  • Peng George Wang
Article

Abstract

The gene galE encoding UDP-galactose 4-epimerase was cloned into E. coli BL21(DE3) from the chromosomal DNA of E. coli strain K-12. High expression of the soluble recombinant epimerase was achieved in the cell lysate. In order to evaluate the use of this epimerase in enzymatic synthesis of important α-Gal epitopes (oligosaccharides with a terminal Galα1,3Gal sequence), a new radioactivity assay (α1,3-galactosyltransferase coupled assay) was established to characterize its activity in producing UDP-galactose from UDP-glucose. Approximately 2700 units (100 mg) enzyme with a specific activity of 27 U mg−1 protein could be obtained from one liter of bacterial culture. The epimerase was active in a wide pH range with an optimum at pH 7.0. This expression system established a viable route to the enzymatic production of α-Gal oligosaccharides to support xenotransplantation research.

cloning E. coli expression α 1,3-galactosyltransferase UDP-galactose 4-epimerase 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Xi Chen
    • 1
  • Przemyslaw Kowal
    • 1
  • Sarah Hamad
    • 1
  • Hongni Fan
    • 1
  • Peng George Wang
    • 1
  1. 1.Department of ChemistryWayne State UniversityDetroitUSA

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