Minireactor-based high-throughput temperature profiling for the optimization of microbial and enzymatic processes

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• 作者：Martin Kunze ; Clemens Lattermann ; Sylvia Diederichs…
• 关键词：High ; throughput screening ; On ; line monitoring ; Microtiter plate ; Optical measurement ; Temperature optimum
• 刊名：Journal of Biological Engineering
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：8
• 期：1
• 全文大小：3,638 KB
• 参考文献：1. Shuler, ML, Kargi, F (2002) Bioprocess engineering : basic concepts. Prentice Hall, Upper Saddle River, NJ
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• 刊物类别：Engineering
• 刊物主题：Biotechnology
  Biomedical Engineering
  Environmental Engineering/Biotechnology
  
• 出版者：BioMed Central
• ISSN：1754-1611

文摘

Background Bioprocesses depend on a number of different operating parameters and temperature is one of the most important ones. Unfortunately, systems for rapid determination of temperature dependent reaction kinetics are rare. Obviously, there is a need for a high-throughput screening procedure of temperature dependent process behavior. Even though, well equipped micro-bioreactors are a promising approach sufficient temperature control is quite challenging and rather complex. Results In this work a unique system is presented combining an optical on-line monitoring device with a customized temperature control unit for 96 well microtiter plates. By exposing microtiter plates to specific temperature profiles, high-throughput temperature optimization for microbial and enzymatic systems in a micro-scale of 200?μL is realized. For single well resolved temperature measurement fluorescence thermometry was used, combining the fluorescent dyes Rhodamin B and Rhodamin 110. The real time monitoring of the microbial and enzymatic reactions provides extensive data output. To evaluate this novel system the temperature optima for Escherichia coli and Kluyveromyces lactis regarding growth and recombinant protein production were determined. Furthermore, the commercial cellulase mixture Celluclast as a representative for enzymes was investigated applying a fluorescent activity assay. Conclusion Microtiter plate-based high-throughput temperature profiling is a convenient tool for characterizing temperature dependent reaction processes. It allows the evaluation of numerous conditions, e.g. microorganisms, enzymes, media, and others, in a short time. The simple temperature control combined with a commercial on-line monitoring device makes it a user friendly system.