背景:小规模的常规实验通常以“黑匣子”的方式进行,仅分析最终样品中的产品浓度。在线监测相关工艺特性和参数,如基板限制,产品抑制和氧气供应不足。因此,迄今为止,对于新的微生物系统的详细研究,需要配备齐全的实验室规模的搅拌罐生物反应器。然而,它们太宽敞了,大量并行操作既费力又昂贵。因此,这项研究的目的是提出一种新的实验方法,通过并行使用两个具有在线监测功能的小规模培养系统来获得密集的定量过程信息:呼吸活性监测系统(RAMOS)和BioLector设备。
结果:将相同的“主混合物”(培养基加微生物)分配到不同的小规模培养系统:1)RAMOS装置;2)用于BioLector装置的48孔微量滴定板;和3)单独的摇瓶或微量滴定板用于离线取样。通过调整相同的最大氧转移能力(OTRmax),RAMOS和BioLector在线监测系统的结果对所有研究的微生物系统都很好地相互补充(E.大肠杆菌G.氧化丹,K.乳酸)和培养条件(氧限制,双氢生长,自动感应,缓冲效果)。
结论:RAMOS和BioLector设备的并行使用是获得有关被评估微生物的生长和生产行为的全面定量数据的合适且快速的方法。这些获得的数据大大减少了实验室规模的搅拌釜生物反应器中用于基本工艺开发的必要实验数量。因此,在更短的时间内并行获得更多的定量信息。
BACKGROUND: Conventional experiments in small scale are often performed in a \'Black Box\' fashion, analyzing only the product concentration in the final sample. Online monitoring of relevant process characteristics and parameters such as substrate limitation, product inhibition and oxygen supply is lacking. Therefore, fully equipped laboratory-scale stirred tank bioreactors are hitherto required for detailed studies of new microbial systems. However, they are too spacious, laborious and expensive to be operated in larger number in parallel. Thus, the aim of this study is to present a new experimental approach to obtain dense quantitative process information by parallel use of two small-scale culture systems with online monitoring capabilities: Respiration Activity MOnitoring System (RAMOS) and the BioLector device.
RESULTS: The same \'mastermix\' (medium plus microorganisms) was distributed to the different small-scale culture systems: 1) RAMOS device; 2) 48-well microtiter plate for BioLector device; and 3) separate shake flasks or microtiter plates for offline sampling. By adjusting the same maximum oxygen transfer capacity (OTRmax), the results from the RAMOS and BioLector online monitoring systems supplemented each other very well for all studied microbial systems (E. coli, G. oxydans, K. lactis) and culture conditions (oxygen limitation, diauxic growth, auto-induction, buffer effects).
CONCLUSIONS: The parallel use of RAMOS and BioLector devices is a suitable and fast approach to gain comprehensive quantitative data about growth and production behavior of the evaluated microorganisms. These acquired data largely reduce the necessary number of experiments in laboratory-scale stirred tank bioreactors for basic process development. Thus, much more quantitative information is obtained in parallel in shorter time.