Cloning and expression of β1,4-galactosyltransferase gene from Helicobacter pylori

被引:25
作者
Endo, T [1 ]
Koizumi, S [1 ]
Tabata, K [1 ]
Ozaki, A [1 ]
机构
[1] Kyowa Hakko Kogyo Co Ltd, Tokyo Res Labs, Tokyo 1948533, Japan
关键词
enzymatic synthesis; glycosyltransferase; oligosaccharide; Helicobacter pylori;
D O I
10.1093/glycob/10.8.809
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Helicobacter pylori, which is a human pathogen associated with gastric and duodenal ulcer, has been shown to express human oncofetal antigens Lewis X and Lewis Y, Although the mammalian glycosyltransferases that synthesize these structures are well characterized, little is known about the corresponding bacterial enzymes. We report that a novel beta 1,4-galactosyltransferase gene (HpgalT) involved in the biosynthesis of lipopolysaccharides in H.pylori has been cloned and expressed in Escherichia coli, The deduced amino acid sequence of the protein (HpGal-T) encoded by HpgalT consists of 274 residues with the calculated molecular mass of 31,731 Da, which does not show significant similarity to those of beta 1,4-galactosyltransferases from mammalian sources and Neisseria. It was confirmed that HpGal-T catalyzed the introduction of galactose from UI)P-Gal in a beta 1,4 linkage to accepting N-acetylglucosamine (GlcNAc) residues by means of high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). When the E.coli cells which overexpressed HpgalT was coupled with the UDP-Gal production system, which consisted of recombinant E.coli cells overexpressing its UDP-Gal biosynthetic genes and Corynebacterium ammoniagenes, N-acetyllactosamine, a core structure of lipopolysaccharide of H.pylori, was efficiently produced from orotic acid, galactose, and GlcNAc.
引用
收藏
页码:809 / 813
页数:5
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