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Production of α‐Galactosyl Epitopes via Combined Use of Two Recombinant Whole Cells Harboring UDP‐Galactose 4‐Epimerase and α‐1,3‐Galactosyltransferase
Author(s) -
Chen Xi,
Zhang Wei,
Wang Jianqiang,
Fang Jianwen,
Wang Peng George
Publication year - 2000
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp000052s
Subject(s) - galactosyltransferase , recombinant dna , epitope , glycosyltransferase , biochemistry , galactose , escherichia coli , lactose , enzyme , beta galactosidase , biology , chemistry , microbiology and biotechnology , gene , antibody , genetics
α‐Galactosyl epitopes (or α‐Gal, oligosaccharides with a terminal Galα1,3Gal sequence) are a class of biologically important oligosaccharides in great demand in bulk quantities for basic and clinical studies on preventing hyperacute rejection in pig‐to‐primate organ xenotransplantaion. A truncated bovine α‐1,3‐galactosyltransferase, the key enzyme responsible for the biosynthesis of the terminal structure of α‐Gal, was cloned and overexpressed previously. The acceptor specificity was further studied in the present paper, and lactose and galactose derivatives were found to be good acceptors. To develop a more proficient reaction process, we report herein an example of an efficient enzymatic synthesis of α‐Gal oligosaccharides catalyzed by the combination of two recombinant Escherichia coli whole cells harboring the genes of a UDP‐galactose 4‐epimerase and the α‐1,3‐galactosyltransferase, respectively. Using lactosyl azide (LacN 3 ) as the acceptor for the glycosyltransferase, the combined use of the two recombinant cells efficiently produced α‐Gal epitope Galα1,3LacN 3 in 60−68% yield.