Highly efficient chemoenzymatic synthesis of α-galactosyl epitopes with a recombinant α(1→3)-galactosyltransferase

Jianwen Fang, Jun Li, Xi Chen, Yingnan Zhang, Jianqiang Wang, Zhengmao Guo, Wei Zhang, Libing Yu, Keith Brew, Peng George Wang

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

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 (α13-GalT) for the synthesis of xenoactive α-galactosyl epitopes, which are highly desired in the research of xenotransp]antation 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.

Original languageEnglish (US)
Pages (from-to)6635-6638
Number of pages4
JournalJournal of the American Chemical Society
Volume120
Issue number27
DOIs
StatePublished - Jul 15 1998
Externally publishedYes

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Galactosyltransferases
Epitopes
Antibodies
Bearings (structural)
Enzymes
Heterologous Transplantation
Carbohydrates
Immunotherapy
Escherichia coli
Regeneration
Anti-Idiotypic Antibodies
Animals
Cells
Serum
Research

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Highly efficient chemoenzymatic synthesis of α-galactosyl epitopes with a recombinant α(1→3)-galactosyltransferase. / Fang, Jianwen; Li, Jun; Chen, Xi; Zhang, Yingnan; Wang, Jianqiang; Guo, Zhengmao; Zhang, Wei; Yu, Libing; Brew, Keith; Wang, Peng George.

In: Journal of the American Chemical Society, Vol. 120, No. 27, 15.07.1998, p. 6635-6638.

Research output: Contribution to journalArticle

Fang, Jianwen ; Li, Jun ; Chen, Xi ; Zhang, Yingnan ; Wang, Jianqiang ; Guo, Zhengmao ; Zhang, Wei ; Yu, Libing ; Brew, Keith ; Wang, Peng George. / Highly efficient chemoenzymatic synthesis of α-galactosyl epitopes with a recombinant α(1→3)-galactosyltransferase. In: Journal of the American Chemical Society. 1998 ; Vol. 120, No. 27. pp. 6635-6638.
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AU - Wang, Jianqiang

AU - Guo, Zhengmao

AU - Zhang, Wei

AU - Yu, Libing

AU - Brew, Keith

AU - Wang, Peng George

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N2 - 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 (α13-GalT) for the synthesis of xenoactive α-galactosyl epitopes, which are highly desired in the research of xenotransp]antation 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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