Miniature bioreactors for automated high-throughput bioprocess design (HTBD): reproducibility of parallel fed-batch cultivations with Escherichia coli.
To verify the reproducibility of cultivations of Escherichia coli in novel millilitre-scale bioreactors, fully automated fed-batch cultivation was performed in seven parallel-operated ml-scale bioreactors with an initial volume of 10 ml/reactor. The process was automatically controlled by a liquid-handling system responsible for glucose feeding, titration and sampling. Atline analysis (carried out externally of the reaction vessel with a short time delay) comprised automated pH and attenuance measurements. The partial pressure of oxygen (pO2) was measured online by a novel fluorimetric sensor block measuring the fluorescence lifetime of fluorophors immobilized inside the millilitre-scale bioreactors. Within a process time of 14.6 h, the parallel cultivation yielded a dry cell weight of 36.9+/-0.9 g.l(-1). Atline pH measurements were characterized by an S.D. of <1.1% throughout the process. Computational-fluid-dynamics simulation of single-phase flow yields a mean power input of 21.9 W.l(-1) at an impeller speed of 2800 rev./min corresponding to a power number (NP) of 3.7.
Puskeiler
Robert
R
Lehrstuhl für Bioverfahrenstechnik, Technische Universität München, Boltzmannstrasse 15, 85748 Garching, Germany. r.puskeiler@lrz.tum.de
Kusterer
Andreas
A
John
Gernot T
GT
Weuster-Botz
Dirk
D
D001331:Automation; D018533:Biomass; D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D003198:Computer Simulation; D003470:Culture Media / Q000737:chemistry; D004867:Equipment Design; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development*; D005285:Fermentation; D005470:Fluorometry; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors
D003470:Culture Media; D005947:Glucose; D010100:Oxygen