Diffusion-driven fed-batch fermentation in perforated ring flasks

Biotechnology Letters, vol. 46, pp. 571–582

Abstract

Abstract Purpose Simultaneous membrane-based feeding and monitoring of the oxygen transfer rate shall be introduced to the newly established perforated ring flask, which consists of a cylindrical glass flask with an additional perforated inner glass ring, for rapid bioprocess development. Methods A 3D-printed adapter was constructed to enable monitoring of the oxygen transfer rate in the perforated ring flasks. Escherichia coli experiments in batch were performed to validate the adapter. Fed-batch experiments with different diffusion rates and feed solutions were performed. Results The adapter and the performed experiments allowed a direct comparison of the perforated ring flasks with Erlenmeyer flasks. In batch cultivations, maximum oxygen transfer capacities of 80 mmol L−1 h−1 were reached with perforated ring flasks, corresponding to a 3.5 times higher capacity than in Erlenmeyer flasks. Fed-batch experiments with a feed reservoir concentration of 500 g glucose L−1 were successfully conducted. Based on the oxygen transfer rate, an ammonium limitation could be observed. By adding 40 g ammonium sulfate L−1 to the feed reservoir, the limitation could be prevented. Conclusion The membrane-based feeding, an online monitoring technique, and the perforated ring flask were successfully combined and offer a new and promising tool for screening and process development in biotechnology.

Authors 5

1. Clara Lüchtrath Aachen AVT‑Biochemical Engineering

   RWTH Aachen University

   Affiliation as printed

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany

2. Felix Lamping Aachen AVT‑Biochemical Engineering

   RWTH Aachen University

   Affiliation as printed

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany

3. Maurice Finger Aachen AVT‑Biochemical Engineering

   RWTH Aachen University

   Affiliation as printed

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany

4. Jørgen Barsett Magnus Aachen AVT‑Biochemical Engineering

   RWTH Aachen University

   Affiliation as printed

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany

5. Jochen Büchs corresponding Aachen AVT‑Biochemical Engineering

   RWTH Aachen University

   Affiliation as printed

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany. jochen.buechs@avt.rwth-aachen.de

   AVT-Biochemical Engineering, RWTH Aachen University, Forckenbeckstraße 51, 52074, Aachen, Germany