Flitsch David, Ladner Tobias, Lukacs Mihaly, Büchs Jochen
Biochemical Engineering, AVT-Aachener Verfahrenstechnik, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.
Microb Cell Fact. 2016 Feb 24;15:45. doi: 10.1186/s12934-016-0444-4.
Despite the progressive miniaturization of bioreactors for screening purposes, shake flasks are still widespread in biotechnological laboratories and industry as cultivation vessels. Shake flasks are often applied as the first or second step in applications such as strain screening or media optimization. Thus, there are ongoing efforts to develop online measurement techniques for shake flasks, to gain as much information as possible about the cultured microbial system. Since dissolved oxygen tension (DOT) is a key experimental parameter, its accurate determination during the course of experiment is critical. Some of the available DOT measurement techniques can lead to erroneous measurements or are very difficult to handle. A reliable and easy to use DOT measurement system, based on suspended oxygen-sensitive nanoparticles, is presented in this work.
In a cultivation of Kluyveromyces lactis, a new DOT measurement technique via suspended oxygen-sensitive nanoparticles was compared with the conventional DOT measurement via fixed sensor spots. These experiments revealed the main disadvantage of applying sensor spots. With further cultivations of Escherichia coli and Hansenula polymorpha, the new measurement technique was successfully validated. In combination with a RAMOS device, kLa values were determined during the presented cultivations. The determined kLa values are in good agreement with a correlation recently found in the literature.
The presented DOT measurement technique via suspended oxygen-sensitive nanoparticles in shake flasks turned out to be easy to use, robust and reliable under all applied combinations of shaking frequencies and filling volumes. Its applicability as an online monitoring system for cultivations was shown by means of four examples. Additionally, in combination with a RAMOS device, the possibility of experimental kLa determination was successfully demonstrated.
尽管用于筛选目的的生物反应器在不断小型化，但摇瓶作为培养容器在生物技术实验室和工业中仍广泛使用。摇瓶常用于菌株筛选或培养基优化等应用的第一步或第二步。因此，人们一直在努力开发用于摇瓶的在线测量技术，以获取关于培养微生物系统的尽可能多的信息。由于溶解氧张力（DOT）是一个关键实验参数，在实验过程中准确测定它至关重要。一些现有的DOT测量技术可能导致测量误差或操作非常困难。本文介绍了一种基于悬浮氧敏感纳米颗粒的可靠且易于使用的DOT测量系统。
在乳酸克鲁维酵母的培养中，将通过悬浮氧敏感纳米颗粒的新型DOT测量技术与通过固定传感器点的传统DOT测量进行了比较。这些实验揭示了应用传感器点的主要缺点。通过大肠杆菌和多形汉逊酵母的进一步培养，成功验证了这种新的测量技术。结合RAMOS装置，在所示培养过程中测定了kLa值。所测定的kLa值与最近文献中发现的相关性良好一致。
本文介绍的摇瓶中通过悬浮氧敏感纳米颗粒的DOT测量技术在所有应用的振荡频率和填充体积组合下都易于使用、稳健且可靠。通过四个例子展示了其作为培养在线监测系统的适用性。此外，结合RAMOS装置，成功证明了实验测定kLa的可能性。
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