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.
Original language | English (US) |
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Pages (from-to) | 1131-1135 |
Number of pages | 5 |
Journal | Biotechnology Letters |
Volume | 21 |
Issue number | 12 |
DOIs | |
State | Published - 1999 |
Externally published | Yes |
Keywords
- α 1,3-galactosyltransferase
- Cloning
- E. coli
- Expression
- UDP-galactose 4- epimerase
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
- Microbiology