Laboratory of Bioprocess Technology, Institute of Biotechnology, Technische Universität Berlin, Ackerstrasse 71-76, ACK 24, 13355 Berlin, Germany.
J Ind Microbiol Biotechnol. 2010 Sep;37(9):935-42. doi: 10.1007/s10295-010-0742-5. Epub 2010 May 29.
The time-dependent development of cell polarizability and length in Escherichia coli batch fermentations were observed at-line with electrooptical measurements. While using a measurement system with fully automated sample preparation, the development of these properties can be observed with a comparable high frequency (six measurements per hour). The polarizability as well as the mean cell length both increase soon after inoculation and then decline from the growth phase on until the stationary phase is reached. Based on the dynamic behavior of polarizability, the growth phase can be divided into four distinct stages. Changes in the cultivation temperature or the pre-cultivation conditions lead to alterations in the development of the polarizability and mean cell length. Based on the frequency disperse of polarizability measured at four different frequencies from 210 to 2,100 kHz, a prediction model is established that is based on the relation of the polarizability to the metabolic activity. Applying multi-linear partial least squares methods (N-PLS), the model is able to predict the specific acetate synthesis and uptake with a root mean square error of prediction of 0.19 (6% of the mean). The method represents a tool for characterization of different stages with respect to microbial metabolic activity and the energy balance during batch cultivations.
采用光电测量法在线观察了大肠杆菌分批发酵中细胞极化率和长度随时间的变化。在使用具有全自动样品制备的测量系统时,可以以相当高的频率(每小时六次测量)观察这些特性的发展。极化率和平均细胞长度在接种后很快增加,然后从生长阶段下降,直到达到静止阶段。基于极化率的动态行为,可以将生长阶段分为四个不同的阶段。培养温度或预培养条件的变化会导致极化率和平均细胞长度的发展发生变化。基于在 210 至 2,100 kHz 的四个不同频率下测量的极化率频散,建立了一个基于极化率与代谢活性关系的预测模型。应用多线性偏最小二乘法(N-PLS),该模型能够以 0.19 的预测均方根误差(平均的 6%)预测特定的乙酸盐合成和吸收。该方法代表了一种用于在分批培养过程中针对微生物代谢活性和能量平衡对不同阶段进行特征描述的工具。