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Low-Frequency Electrochemical Impedance Spectroscopy as a Monitoring Tool for Yeast Growth in Industrial Brewing Processes

机译:低频电化学阻抗谱作为工业酿造过程中酵母生长的监测工具

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摘要

Today’s yeast total biomass and viability measurements during the brewing process are dependent on offline methods such as methylene blue or florescence dye-based staining, and/or the usage of flow cytometric measurements. Additionally, microscopic cell counting methods decelerate an easy and quick prediction of yeast viability. These processes are time consuming and result in a time-delayed response signal, which not only reduces the knowledge of the performance of the yeast itself, but also impacts the quality of the final product. Novel approaches in process monitoring during the aerobic and anaerobic fermentation of Saccharomyces cerevisiae are not only limited to classical pH, dO 2 and off-gas analysis, but they also use different in situ and online sensors based on different physical principles to determine the biomass, product quality and cell death. Within this contribution, electrochemical impedance spectroscopy (EIS) was used to monitor the biomass produced in aerobic and anaerobic batch cultivation approaches, simulating the propagation and fermentation unit operation of industrial brewing processes. Increases in the double-layer capacitance (C DL ), determined at frequencies below 1 kHz, were proportional to the increase of biomass in the batch, which was monitored in the online and inline mode. A good correlation of C DL with the cell density was found. In order to prove the robustness and flexibility of this novel method, different state-of-the-art biomass measurements (dry cell weight—DCW and optical density—OD) were performed for comparison. Because measurements in this frequency range are largely determined by the double-layer region between the electrode and media, rather minor interferences with process parameters (aeration and stirring) were to be expected. It is shown that impedance spectroscopy at low frequencies is not only a powerful tool for the monitoring of viable yeast cell concentrations during operation, but it is also perfectly suited to determining physiological states of the cells, and may facilitate biomass monitoring in the brewing and yeast-propagating industry drastically.
机译:酿造过程中,当今酵母的总生物量和生存能力的测量取决于离线方法,例如基于亚甲基蓝或荧光染料的染色,和/或流式细胞仪的使用。另外,显微细胞计数方法降低了对酵母活力的容易和快速的预测。这些过程很耗时并且导致时间延迟的响应信号,这不仅降低了酵母本身性能的知识,而且还影响了最终产品的质量。酿酒酵母需氧发酵和厌氧发酵过程监控的新方法不仅限于经典pH,dO 2和废气分析,而且还基于不同的物理原理使用不同的原位和在线传感器来确定生物量,产品质量和细胞死亡。在此贡献范围内,电化学阻抗谱(EIS)用于监测有氧和厌氧分批培养方法中产生的生物量,模拟工业酿造过程的繁殖和发酵单元操作。在低于1 kHz的频率下确定的双层电容(C DL)的增加与批次中生物量的增加成比例,可以在线和在线方式对其进行监控。发现C DL与细胞密度具有良好的相关性。为了证明这种新方法的鲁棒性和灵活性,进行了不同水平的生物量测量(干细胞重量-DCW和光密度-OD)进行比较。由于在此频率范围内的测量很大程度上取决于电极和介质之间的双层区域,因此,预计会对工艺参数(充气和搅拌)产生较小的干扰。结果表明,低频阻抗谱不仅是操作过程中监测活酵母细胞浓度的有力工具,而且还非常适合确定细胞的生理状态,并可能有助于酿造和酵母菌中生物量的监测。大力传播行业。

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