Controlling pH in shake flasks using polymer-based controlled-release discs with pre-determined release kinetics

BMC Biotechnology

Research Article

Hideo Ikeda¹  Johannes Klinger¹  Marco Scheidle¹  Jochen Büchs¹  Doris Klee²  Barbara Dittrich² 

[1] Aachener Verfahrenstechnik - Biochemical Engineering, RWTH Aachen University, Sammelbau Biologie, Worringer Weg 1, D-52074, Aachen, Germany;Textile Chemistry and Macromolecular Chemistry, RWTH Aachen University, Pauwelsstr. 8, 52074, Aachen, Germany;

关键词: Shake Flask;    Sodium Carbonate;    Release Kinetic;    Buffer Concentration;    Oxygen Transfer Rate;   

DOI  :  10.1186/1472-6750-11-25

 received in 2010-07-21, accepted in 2011-03-23,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThere are significant differences in the culture conditions between small-scale screenings and large-scale fermentation processes. Production processes are usually conducted in fed-batch cultivation mode with active pH-monitoring and control. In contrast, screening experiments in shake flasks are usually conducted in batch mode without active pH-control, but with high buffer concentrations to prevent excessive pH-drifts. These differences make it difficult to compare results from screening experiments and laboratory and technical scale cultivations and, thus, complicate rational process development. In particular, the pH-value plays an important role in fermentation processes due to the narrow physiological or optimal pH-range of microorganisms. To reduce the differences between the scales and to establish a pH-control in shake flasks, a newly developed easy to use polymer-based controlled-release system is presented in this paper. This system consists of bio-compatible silicone discs embedding the alkaline reagent Na2CO3. Since the sodium carbonate is gradually released from the discs in pre-determined kinetics, it will ultimately compensate the decrease in pH caused by the biological activity of microorganisms.ResultsThe controlled-release discs presented here were successfully used to cultivate E. coli K12 and E. coli BL21 pRSET eYFP-IL6 in mineral media with glucose and glycerol as carbon (C) sources, respectively. With glucose as the C-source it was possible to reduce the required buffer concentration in shake flask cultures by 50%. Moreover, with glycerol as the C-source, no buffer was needed at all.ConclusionsThese novel polymer-based controlled-release discs allowed buffer concentrations in shake flask media to be substantially reduced or omitted, while the pH remains in the physiological range of the microorganisms during the whole cultivation time. Therefore, the controlled-release discs allow a better control of the pH, than merely using high buffer concentrations. The conditions applied here, i.e. with significantly reduced buffer concentrations, enhance the comparability of the culture conditions used in screening experiments and large-scale fermentation processes.

【 授权许可】

Unknown   
© Scheidle et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.