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Parallelized microscale fed-batch cultivation in online-monitored microtiter plates: implications of media composition and feed strategies for process design and performance.
Journal of Industrial Microbiology & Biotechnology ( IF 3.2 ) Pub Date : 2019-10-31 , DOI: 10.1007/s10295-019-02243-w
Holger Morschett 1 , Roman Jansen 1, 2 , Christian Neuendorf 1 , Matthias Moch 1 , Wolfgang Wiechert 1, 3 , Marco Oldiges 1, 2
Affiliation  

Limited throughput represents a substantial drawback during bioprocess development. In recent years, several commercial microbioreactor systems have emerged featuring parallelized experimentation with optical monitoring. However, many devices remain limited to batch mode and do not represent the fed-batch strategy typically applied on an industrial scale. A workflow for 32-fold parallelized microscale cultivation of protein secreting Corynebacterium glutamicum in microtiter plates incorporating online monitoring, pH control and feeding was developed and validated. Critical interference of the essential media component protocatechuic acid with pH measurement was revealed, but was effectively resolved by 80% concentration reduction without affecting biological performance. Microfluidic pH control and feeding (pulsed, constant and exponential) were successfully implemented: Whereas pH control improved performance only slightly, feeding revealed a much higher optimization potential. Exponential feeding with µ = 0.1 h-1 resulted in the highest product titers. In contrast, other performance indicators such as biomass-specific or volumetric productivity resulted in different optimal feeding regimes.

中文翻译:


在线监测微量滴定板中的并行微量补料分批培养:培养基组成和补料策略对工艺设计和性能的影响。



有限的通量是生物工艺开发过程中的一个重大缺陷。近年来,出现了几种具有光学监测并行实验特点的商业微生物反应器系统。然而,许多设备仍然仅限于分批模式,并不代表通常应用于工业规模的补料分批策略。开发并验证了在微量滴定板中对分泌蛋白质的谷氨酸棒杆菌进行 32 倍并行微量培养的工作流程,其中包括在线监测、pH 控制和补料。培养基中的主要成分原儿茶酸对 pH 测量存在严重干扰,但通过降低 80% 的浓度得到了有效解决,且不影响生物性能。微流体 pH 控制和进料(脉冲式、恒定式和指数式)已成功实施:虽然 pH 控制仅略微改善了性能,但进料显示出更高的优化潜力。 µ = 0.1 h-1 的指数饲喂可产生最高的产物滴度。相比之下,其他性能指标(例如生物量生产率或体积生产率)导致不同的最佳饲喂方案。
更新日期:2020-03-07
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