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Impact of oscillating conditions on the synthesis and incorporation of branched-chain amino acids in Escherichia coli



Profound changes in the amino acid pool can be detected when an E. coli culture is opposed to oscillating conditions. This likely influences the composition of recombinant proteins. It was found that the synthesis of norvaline in Escherichia coli is strongly accelerated under oscillating conditions. The changed dynamics of the (branched-chain) amino acids’ synthesis is quantified by a dynamic flux analysis and compared to non-oscillating conditions as they occur in homogeneous lab-scale bioreactors. Short term responses are observed by combining the scale-down reactor and a rapid sampling unit Bioscope. The composition of amino acids in the protein fraction is observed with mass spectrometry. The presented methodology should be understood as a bridge between systems biology for understanding the regulatory network of a cell and the industrial application for strain and process development.



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