Production of recombinant xenotransplantation antigen in Escherichia coli.
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 Galp alpha(1-3)Galp beta(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), beta(1-4) galactosyltransferase LgtB, and bovine alpha(1-3) galactosyltransferase GstA. This strain produced 0.68 g/L of the heptasaccharide Galp alpha(1-3)Galp beta(1-4)(GlcNAc)(5), which harbours the xenoantigen at its non-reducing end, establishing the feasibility of this approach.
Bettler
Emmanuel
E
Centre de Recherches sur les Macromolécules Végétales (CNRS), Affiliated with Joseph Fourier University, BP 53X, 38041 Grenoble cedex 9, France. e.bettler@cmbi.kun.nl
Imberty
Anne
A
Priem
Bernard
B
Chazalet
Valérie
V
Heyraud
Alain
A
Joziasse
David H
DH
Geremia
Roberto A
RA
D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000378:metabolism; D002417:Cattle; D002845:Chromatography; D000939:Epitopes; D004926:Escherichia coli / Q000276:immunology* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000276:immunology / Q000378:metabolism*; D018014:Gene Transfer Techniques; D016695:Glycosyltransferases / Q000235:genetics*; D047908:Intracellular Signaling Peptides and Proteins; D008954:Models, Biological; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D009237:N-Acetyllactosamine Synthase / Q000276:immunology / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011134:Polysaccharides / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000737:chemistry; D013552:Swine; D013696:Temperature; D013997:Time Factors; D014183:Transplantation, Heterologous / Q000276:immunology*; D014312:Trisaccharides
D000941:Antigens; D001426:Bacterial Proteins; D002352:Carrier Proteins; D000939:Epitopes; D029968:Escherichia coli Proteins; C105760:GstA protein, bacteria; D047908:Intracellular Signaling Peptides and Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D014312:Trisaccharides; C076569:alpha-galactosyl epitope; D016695:Glycosyltransferases; D017351:N-Acetylglucosaminyltransferases; C060771:NodC protein, Rhizobiales; D009237:N-Acetyllactosamine Synthase; C473806:LgtB protein, E coli