Highly Efficient Chemoenzymatic Synthesis of α-Galactosyl Epitopes with a Recombinant α(1→3)-Galactosyltransferase
α-Galactosyl epitopes are carbohydrate structures bearing a Galα1-3Galβ terminus. The interaction of these epitopes on the surface of animal cells with anti-α-galactosyl antibodies in human serum is believed to be the main cause in antibody-mediated hyperacute rejection in xenotransplantation. This...
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Published in | Journal of the American Chemical Society Vol. 120; no. 27; pp. 6635 - 6638 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
WASHINGTON
American Chemical Society
15.07.1998
Amer Chemical Soc |
Subjects | |
Online Access | Get full text |
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Summary: | α-Galactosyl epitopes are carbohydrate structures bearing a Galα1-3Galβ terminus. The interaction of these epitopes on the surface of animal cells with anti-α-galactosyl antibodies in human serum is believed to be the main cause in antibody-mediated hyperacute rejection in xenotransplantation. This report describes an efficient chemoenzymatic approach based on the use of recombinant α(1→3)-galactosyltransferase (α1,3-GalT) for the synthesis of xenoactive α-galactosyl epitopes, which are highly desired in the research of xenotransplantation and immunotherapy. A truncated bovine α1,3-GalT (80−368) was cloned into the pET15b vector and subsequently transformed into E. coli BL21 strain. This expression system efficiently produced the soluble recombinant enzyme on a large scale with highly specific activity. A variety of α(1→3)-galactosylated epitopes were synthesized using such a recombinant enzyme. In a unique fashion, α-galactosyl pentasaccharide was synthesized via a one-pot, two-step enzymatic synthesis with in situ cofactor regeneration. |
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Bibliography: | istex:F5D4C4B46A5ADEADEF78BDD8C5836AD4307B63B4 ark:/67375/TPS-TN2HXGWZ-K |
ISSN: | 0002-7863 1520-5126 |
DOI: | 10.1021/ja9808898 |