Department of Chemistry and Biotechnology, Faculty of Science and Technology, Kochi University, 2-5-1 Akebono-cho, Kochi City, Kochi 780-8520, Japan.
Department of Brewing, Tsukasa Botan Brewing Company, Limited, 1299 Ko, Sakawa-cho, Takaoka-gun, Kochi 789-1201, Japan.
Anal Methods. 2024 Jun 27;16(25):4045-4053. doi: 10.1039/d4ay00591k.
Facile and effective analysis methods are desirable for elucidating the behaviours of metabolites during fermentation reactions. Herein, a multifunctional-separation-mode ion chromatography (MFS-IC) method was developed for the simultaneous monitoring of major metabolites during multiple parallel fermentation, including those related to central carbon metabolism (saccharification, glycolysis, alcoholic fermentation, and the tricarboxylic acid (TCA) cycle). The use of two types of sulfo-modified size-exclusion columns and phthalic acid as the eluent allowed the separation of oligosaccharides (disaccharides, trisaccharides, and tetrasaccharides), glucose, pyruvate, and major organic acids during the TCA cycle (-aconitate, citrate, -citrate, malate, fumarate, and succinate but not α-ketoglutarate) from other non-target analytes. The MFS-IC method was successfully applied to monitoring the major metabolites in the rice wine brewing process. This approach can contribute to an improved understanding of metabolite behaviour during fermentation without requiring the use of expensive advanced instrumentation methods such as liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.
需要简便有效的分析方法来阐明发酵反应过程中代谢物的行为。本文开发了一种多功能分离模式离子色谱(MFS-IC)方法,用于同时监测多个平行发酵过程中的主要代谢物,包括与中心碳代谢(糖化、糖酵解、酒精发酵和三羧酸(TCA)循环)相关的代谢物。使用两种类型的磺化修饰的排阻色谱柱和邻苯二甲酸作为洗脱液,可将寡糖(二糖、三糖和四糖)、葡萄糖、丙酮酸和 TCA 循环中的主要有机酸(延胡索酸、柠檬酸、-柠檬酸盐、苹果酸、富马酸和琥珀酸,但不包括α-酮戊二酸)与其他非目标分析物分离。MFS-IC 方法成功应用于监测米酒酿造过程中的主要代谢物。该方法无需使用昂贵的先进仪器方法(如液相色谱-质谱和气相色谱-质谱),有助于更好地了解发酵过程中的代谢物行为。