应用激光镊子拉曼光谱技术收集500L发酵罐中木薯淀粉浓醪乙醇发酵过程底物、产物及酵母单细胞的拉曼光谱,以期从单细胞水平为乙醇发酵提供新的认识。结果显示:1)拉曼光谱可以实时监测浓醪乙醇发酵过程底物与产物的变化;2)酵母细胞胞内物质的变化存在类似于产物变化的前发酵期、主发酵期和后发酵期3个阶段,但出现的时间要比产物变化晚约4h;3)为适应浓醪发酵环境,酵母细胞的生理状态和胞内物质在不断地进行调整,随着环境乙醇浓度的升高,酵母细胞在胞内累积蛋白质和脂类物质,蛋白质二级结构逐渐变为以无规则卷曲为主;4)发酵后期,酵母细胞在胞内累积大量的嘌呤类物质,但细胞间含量存在异质性。上述结果表明,单细胞拉曼光谱技术提供了一种研究微生物发酵的新方法,可从新的角度获知乙醇发酵过程酵母细胞内外的变化信息。 Laser Raman spectroscopy was used to collect the Raman spectra of substrate, product and single cell of yeast in cassava starch fermentations in a 500L fermentor, in order to provide a new understanding of ethanol fermentation from the single cell level. The results show that: 1) Raman spectroscopy can be used to monitor the changes of substrates and products in the ethanol fermentation process in real time; 2) The change of the intracellular material in the yeast cells is similar to that of the product before the fermentation, the main fermentation and the post-fermentation 3 A period of time, but the time of occurrence is about 4h later than the change of the product; 3) to adapt to the fermentation environment of thick mash, the physiological state of yeast cells and intracellular substances are constantly adjusted, with the environmental ethanol concentration, yeast cells In the intracellular accumulation of protein and lipid substances, protein secondary structure gradually becomes irregular curly; 4) During the fermentation, yeast cells accumulate a large number of purine substances in the cell, but there is heterogeneity in the intercellular content. The above results show that the single-cell Raman spectroscopy provides a new method for the study of microbial fermentation, which can be used to obtain information on the changes of yeast cells both inside and outside the ethanol fermentation process from a new perspective.