Production of recombinant xenotransplantation antigen in Escherichia coli

• Published in: Biochemical and biophysical research communications 2003-03, Vol.302 (3), p.620-624
• Main Authors: Bettler, Emmanuel, Imberty, Anne, Priem, Bernard, Chazalet, Valérie, Heyraud, Alain, Joziasse, David H, Geremia, Roberto A
• Format: Article
• Language: English
• Subjects: Animals; Antigens - chemistry; Antigens - metabolism; Bacterial Proteins; Carbohydrate biotechnology; Carrier Proteins - metabolism; Cattle; Chromatography; Epitopes; Escherichia coli - immunology; Escherichia coli - metabolism; Escherichia coli Proteins - immunology; Escherichia coli Proteins - metabolism; Gene Transfer Techniques; Glycosyltransferases; Glycosyltransferases - genetics; Models, Biological; N-Acetylglucosaminyltransferases - metabolism; N-Acetyllactosamine Synthase - immunology; N-Acetyllactosamine Synthase - metabolism; Oligosaccharide production; Plasmids - metabolism; Polysaccharides - biosynthesis; Recombinant Proteins - chemistry; Swine; Temperature; Time Factors; Transplantation, Heterologous - immunology; Trisaccharides; Xenotransplantation antigen
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/S0006-291X(03)00227-4

The synthesis of sufficient amounts of oligosaccharides is the bottleneck for the study of their biological function and their possible use as drug. As an alternative for chemical synthesis, we propose to use Escherichia coli as a “living factory.” We have addressed the production of the Gal pα(1–3)Gal pβ(1–4)GlcNAc epitope, the major porcine antigen responsible for xenograft rejection. An E. coli strain was generated which simultaneously expresses NodC (to provide the chitin-pentaose acceptor), β(1–4) galactosyltransferase LgtB, and bovine α(1–3) galactosyltransferase GstA. This strain produced 0.68 g/L of the heptasaccharide Gal pα(1–3)Gal pβ(1–4)(GlcNAc) 5, which harbours the xenoantigen at its non-reducing end, establishing the feasibility of this approach.