Invertases are glycosidases applied for synthesis of alkyl glycosides that are important and effective surfactants. Stability of invertases in the environment with increased content of organic solvent is crucial for increase of productivity of glycosidases. Their stability is significantly influenced by N-glycosylation. However, yeast N-glycosylation pathways may synthesize plethora of N-glycan structures. A total natural crude mixture of invertase glycoforms (EINV) extracted from Saccharomyces cerevisiae was subfractionated by anion-exchange chromatography on industrial monolithic supports to obtain different glycoforms (EINV1–EINV3). Separated glycoforms exhibited different stabilities in water-alcohol solutions that are in direct correlation with the amount of phosphate bound to N-glycans. Observed differences in stability of different invertase glycoforms were used to improve productivity of methyl β-d-fructofuranoside (MF) synthesis. The efficiency and yield of MF synthesis were improved more than 50% when the most stabile glycoform bearing the lowest amount of phosphorylated N-glycans is selected and utilized. These data underline the importance of analysis of glycan structures attached to glycoproteins, demonstrate different impact of N-glycans on the surface charge and enzyme stability in regard to particular reaction environment, and provide a platform for improvement of yield of industrial enzymatic synthesis by chromatographic selection of glycoforms on monolithic supports.

中文翻译：

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通过色谱选择酵母转化酶的不同 N-糖型提高酶促合成的产量

转化酶是用于合成烷基糖苷的糖苷酶，烷基糖苷是重要且有效的表面活性剂。随着有机溶剂含量的增加，蔗糖酶在环境中的稳定性对于提高糖苷酶的生产力至关重要。它们的稳定性受N-糖基化的显着影响。然而，酵母N-糖基化途径可以合成过多的N-聚糖结构。从酿酒酵母中提取的转化酶糖型 (EINV) 的总天然粗混合物通过阴离子交换色谱在工业整体载体上进行亚分级分离以获得不同的糖型（EINV1-EINV3）。分离的糖型在水-醇溶液中表现出不同的稳定性，这与与N-聚糖结合的磷酸盐的量直接相关。观察到的不同转化酶糖型的稳定性差异用于提高甲基 β- d-呋喃果糖苷 (MF) 合成的生产率。The efficiency and yield of MF synthesis were improved more than 50% when the most stabile glycoform bearing the lowest amount of phosphorylated N -glycans is selected and utilized. 这些数据强调了分析附着在糖蛋白上的聚糖结构的重要性，证明了N的不同影响-聚糖上的表面电荷和酶在特定反应环境下的稳定性，并为通过色谱选择整体载体上的糖型来提高工业酶促合成的产率提供平台。