Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan.
J Biosci Bioeng. 2020 Aug;130(2):128-136. doi: 10.1016/j.jbiosc.2020.03.008. Epub 2020 Apr 4.
The N- and O-linked oligosaccharides from fission yeast Schizosaccharomyces pombe not only contain large amounts of d-mannose (Man) but also contain large amounts of d-galactose (Gal). Although the galactomannans of S. pombe are mainly composed of α1,2- or α1,3-linked Gals, some of the terminal α1,2-linked Gals are found to be linked to pyruvylated β1,3-linked galactose (PvGal). We have determined the structural characteristics of the N-glycans and O-glycans in three Schizosaccharomyces species (S. japonicus, S. octosporus, and S. cryophilus) using lectin blot, H NMR spectroscopy, and size-fractionation high performance liquid chromatography (HPLC), and found that the galactosylation of oligosaccharides was a common feature in fission yeasts. In addition, each of the terminal Galα1,2-, Galβ1,3- and non-substituted Man residues exhibited distinct characteristics. A BLAST search of gene databases in Schizosaccharomyces identified genes homologous to pvg1 encoding pyruvyltransferase of S. pombe. These genes, when expressed in an S. pombe pvg1Δ strains, led to the pyruvylation of non-reducing terminal β-linked Gal, suggesting the biosynthetic pathway of PvGal-containing oligosaccharides is highly conserved in fission yeasts.
裂殖酵母 Schizosaccharomyces pombe 的 N- 和 O-连接寡糖不仅含有大量的 d-甘露糖 (Man),还含有大量的 d-半乳糖 (Gal)。虽然裂殖酵母 Schizosaccharomyces pombe 的半乳甘露聚糖主要由α1,2-或α1,3-连接的 Gal 组成,但一些末端α1,2-连接的 Gal 被发现与丙酮酸化的β1,3-连接的半乳糖 (PvGal) 相连。我们使用凝集素印迹、H NMR 光谱和分级分离高效液相色谱 (HPLC) 确定了三种裂殖酵母物种 (S. japonicus、S. octosporus 和 S. cryophilus) 中的 N-聚糖和 O-聚糖的结构特征,发现寡糖的半乳糖化是裂殖酵母的共同特征。此外,每个末端 Galα1,2-、Galβ1,3-和未取代的 Man 残基都表现出独特的特征。在裂殖酵母的基因数据库中进行 BLAST 搜索,鉴定出与 S. pombe 的 pvg1 编码丙酮酸转移酶同源的基因。这些基因在 S. pombe pvg1Δ 菌株中表达时,导致非还原末端β-连接 Gal 的丙酮酸化,表明含有 PvGal 的寡糖的生物合成途径在裂殖酵母中高度保守。