ISSN:
1573-6776
Keywords:
cloning
;
E. coli
;
expression
;
α 1,3-galactosyltransferase
;
UDP-galactose 4-epimerase
Source:
Springer Online Journal Archives 1860-2000
Topics:
Process Engineering, Biotechnology, Nutrition Technology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1005678225031
Permalink
