[en] Knowledge of concentrations of intracellular metabolites is important for quantitative analysis of metabolic networks. As far as the very fast response of intracellular metabolites in the millisecond range is concerned, the frequently used pulse technique shows an inherent limitation. The time span between the disturbance and the first sample is constrained by the time necessary to obtain a homogeneous distribution of the pertubation within the bioreactor. For determination of rapid changes, a novel sampling technique based on the stopped-flow method has been developed. A continuous stream of biosuspension leaving the bioreactor is being mixed with a glucose solution in a turbulent mixing chamber. Through computer-aided activation of sequentially positioned three-way valves, different residence times and thus reaction times can be verified. The application of this new sampling method is illustrated with examples including measurements of adenine nucleotides and glucose-6-phosphate in Saccharomyces cerevisiae as well as measurements related to the PTS system in Escherichia coli.
Disciplines :
Biotechnology
Author, co-author :
Buziol, Stefan; Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany
Bashir, Imtiaz; Institute of Food Technology, Department of Biotechnology, University of Hohenheim, D-70599 Stuttgart, Germany
Baumeister, Anja; Institute of Food Technology, Department of Biotechnology, University of Hohenheim, D-70599 Stuttgart, Germany
Claassen, Wolfgang; Institute of Food Technology, Department of Biotechnology, University of Hohenheim, D-70599 Stuttgart, Germany
Noisommit-Rizzi, Naruemol; Institute of Biochemical Engineering, University of Stuttgart, D-70569 Stuttgart, Germany
SAUTER, Thomas ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
Mailinger, Werner; Institute of Biochemical Engineering, University of Stuttgart, D-70569 Stuttgart, Germany
Reuss, Matthias
External co-authors :
yes
Language :
English
Title :
New bioreactor-coupled rapid stopped-flow sampling technique for measurements of metabolite dynamics on a subsecond time scale.
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