Assunção Bicca Saul, Sieczkowski Nathalie, Schneider Rémi, Mekoue Julie, Poncet-Legrand Céline, Doco Thierry
Unité Mixte de Recherche Sciences Pour l'Œnologie, Institut Agro, INRAE, Université de Montpellier, Montpellier, France.
Lallemand, SAS, 19 rue des Briquetiers, BP 59, 31702 Blagnac, France.
Carbohydr Polym. 2025 Sep 1;363:123770. doi: 10.1016/j.carbpol.2025.123770. Epub 2025 May 16.
A thorough characterization of the polysaccharide part of Mannoprotein Pools (MPs) from nine different yeast strains was performed to assess biosynthesis-pathways impact on mannoproteins macromolecular structure and later establish structure-function relationships regarding stabilization properties. The strains comprised one wild-type S. cerevisiae strain, its mutants ΔMnn2 and ΔMnn4 (non-ramified N-glycosylated backbone and lacking mannosyl-phosphorylation, respectively), enological Saccharomyces cerevisiae strains grown in different metabolic conditions (aerobic and anaerobic), and three non-Saccharomyces species: Hanseniaspora vineae, Torulaspora delbrueckii, and Schizosaccharomyces japonicus. Neutral sugars composition for MPs of all strains had mainly mannose but Schizosaccharomyces japonicus strain also presented approximately 33 % of galactose. MPs from non-Saccharomyces strains exhibited significant differences in glycosyl-linkage analysis, especially those from Torulaspora delbrueckii and S. japonicus strains. Global net charge analysis of MP from S. japonicus strain suggests the presence of unknown negatively charged ions. These results dismiss the general N-glycosylated and O-Mannosylated structures obtained from S. cerevisiae. HPSEC-MALLS-QELS-Viscometry analysis revealed higher intrinsic viscosity profiles ([η]xMw) of H. vineae and S. japonicus MPs than S. cerevisiae ones. Metabolic conditions also influenced S. cerevisiae MPs biosynthesis: aerobiosis produced more negatively charged (20C.g, net charge at pH 3.5) and less compact (ρ = 2.8) MPs than anaerobiosis (5C.g and 1.4).
对来自九种不同酵母菌株的甘露糖蛋白池(MPs)的多糖部分进行了全面表征,以评估生物合成途径对甘露糖蛋白大分子结构的影响,并随后建立关于稳定特性的结构-功能关系。这些菌株包括一株野生型酿酒酵母菌株、其突变体ΔMnn2和ΔMnn4(分别为无分支的N-糖基化主链和缺乏甘露糖基磷酸化)、在不同代谢条件(有氧和厌氧)下生长的酿酒酵母酿酒菌株,以及三种非酿酒酵母物种:葡萄汉逊酵母、德氏有孢圆酵母和日本裂殖酵母。所有菌株的MPs的中性糖组成主要为甘露糖,但日本裂殖酵母菌株还含有约33%的半乳糖。来自非酿酒酵母菌株的MPs在糖基连接分析中表现出显著差异,尤其是来自德氏有孢圆酵母和日本裂殖酵母菌株的MPs。对日本裂殖酵母菌株的MP的整体净电荷分析表明存在未知的带负电荷离子。这些结果否定了从酿酒酵母获得的一般N-糖基化和O-甘露糖基化结构。高效体积排阻色谱-多角度激光光散射-准弹性光散射-粘度测定分析表明,葡萄汉逊酵母和日本裂殖酵母的MPs的特性粘度谱([η]xMw)高于酿酒酵母的MPs。代谢条件也影响酿酒酵母MPs的生物合成:需氧条件下产生的MPs带负电荷更多(20C.g,pH 3.5时的净电荷)且结构更松散(ρ = 2.8),而厌氧条件下产生的MPs带负电荷较少(5C.g和1.4)。