Highly Efficient Chemoenzymatic Synthesis of -Galactosyl Epitopes with a Recombinant 详细信息    查看全文

• 作者：Jianwen Fang ; Jun Li ; Xi Chen ; Yingnan Zhang ; Jianqiang Wang ; Zhengmao Guo ; Wei Zhang ; Libing Yu ; Keith Brew ; and Peng George Wang
• 刊名：Journal of the American Chemical Society
• 出版年：1998
• 出版时间：July 15, 1998
• 年：1998
• 卷：120
• 期：27
• 页码：6635 - 6638
• 全文大小：154K
• 年卷期：v.120,no.27(July 15, 1998)
• ISSN：1520-5126

文摘

-Galactosyl epitopes are carbohydrate structures bearing a Gal1-3Gal terminus. The interactionof these epitopes on the surface of animal cells with anti--galactosyl antibodies in human serum is believedto be the main cause in antibody-mediated hyperacute rejection in xenotransplantation. This report describesan efficient chemoenzymatic approach based on the use of recombinant (13)-galactosyltransferase (1,3-GalT) for the synthesis of xenoactive -galactosyl epitopes, which are highly desired in the research ofxenotransplantation and immunotherapy. A truncated bovine 1,3-GalT (80-368) was cloned into the pET15bvector and subsequently transformed into E. coli BL21 strain. This expression system efficiently producedthe soluble recombinant enzyme on a large scale with highly specific activity. A variety of (13)-galactosylated epitopes were synthesized using such a recombinant enzyme. In a unique fashion, -galactosylpentasaccharide was synthesized via a one-pot, two-step enzymatic synthesis with in situ cofactor regeneration.