On‐Line Measurement of Culture Fluorescence for Process Monitoring and Control of Biotechnological Processes
On‐Line Measurement of Culture Fluorescence for Process Monitoring and Control of Biotechnological Processes
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在线测量培养物荧光，用于生物技术过程的过程监测和控制
DOI：
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发表时间：
1987
影响因子：
5.2
通讯作者：
K. Schügerl
中科院分区：
文献类型：
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作者：
T. Scheper;T. Lorenz;W. Schmidt;K. Schügerl
在生物反应器中，代谢活跃的细胞被不断变化的化学和物理环境所包围。为了控制这样一个复杂的系统，必须知道细胞与其环境之间的关系。为此，关于细胞内以及细胞与其环境之间的调节过程的信息是重要的。必须连续监测细胞特性、介质组成和物理环境（特别是多相系统的流体动力学）。不幸的是，仍然缺乏用于在培养期间连续监测细胞特性的仪器和传感器。提供细胞代谢状态信息的唯一连续监测技术是使用微型荧光计探针的原位荧光测定法。1957年，Duysens和Amesz首次报道了在活微生物中测量一种关键细胞代谢产物-核苷酸NAD（P）H。为此目的，可以用UV光照射细胞培养物，并且可以检测NAD（P）H荧光。1970年，Harrison和Chance首次使用一种特殊的荧光计来监测生物技术过程。在接下来的几年里，只有少数作者报道了这种测量技术在生物技术中的应用。1981年，Beyeler开发了一种微型荧光传感器，8在接下来的几年中，对这种技术的兴趣越来越大，NAD（P）H依赖性培养物荧光的在线测量可以直接洞察培养过程中的微生物。通过在培养基中添加特殊的示踪剂，荧光测定法还可用于研究生物反应器中的混合时间行为。为了研究混合时间实验过程中的培养物荧光，构建了一种具有特殊光纤的小型化荧光计，用于同时检测两个不同的波长，并将其应用于过程监控和混合时间实验。此外，研究了用于测量培养物荧光（运动发酵单胞菌）的市售显微荧光计。
In bioreactors, metabolically active cells are surrounded by a changing chemical and physical environment. To control such a complex system, the relationship between cells and their environment must be known. For this purpose, information on the regulation process within the cells and between the cells and their environment is important. The cell characteristics, the medium composition, and the physical environment (especially the fluid dynamics of the multiphase system) have to be monitored continuously. Unfortunately, there is still a lack in instrumentation and sensors for continuous monitoring of cell properties during cultivation. The only continuous monitoring technique providing information on the metabolic state of the cells is in situ fluorometry with a miniaturized fluorometer probe. In 1957, Duysens and Amesz first reported on the measurement of one of the key cell metabolites-the nucleotide NAD(P)H-in living microorganisms.’ For this purpose, cell cultures can be irradiated with UV-light, and the NAD(P)H fluorescence light can be detected. In 1970, a special fluorometer was used by Harrison and Chance for the first time to monitor biotechnological processes.’ During the following years, only a few authors reported about applications of this measuring technique in biotechn~logy.~’ In 198 1, Beyeler developed a miniaturized fluorosensor,8 and during the next few years, the interest in this technique increased more and The on-line measurement of NAD(P)H-dependent culture fluorescence gives a direct insight into the microorganisms during cultivation. By adding special tracers to the cultivation medium, fluorometry can also be used for studying the mixing-time behavior in bioreactors. In order to study the culture fluorescence during mixing-time experiments, a miniaturized fluorometer with a special fiber optics for the simultaneous detection of two different wavelengths was constructed and applied to process monitoring and mixing-time experiments. In addition, a commercially available microfluorometer was studied for the measurements of culture fluorescence (Zymomonas mobilis) .