Accumulation of amino acids deriving from pyruvate in Escherichia coli W3110 during fed-batch cultivation in a two-compartment scale-down bioreactor

Bioprocess scale-down simulators are used to investigate the phenomena occurring in industrial scale cultivations. The aim is to simulate the large-scale conditions, characterized by long mixing times introducing local gradients into the bioreactor, in laboratory scale bioreactor. We have expanded the current physiological knowledge about the consequences of such cultivation conditions from well-known mixed-acid fermentation to amino acid metabolism. Several interesting deviations were observed for amino acids derived from pyruvate when a STR-PFR scale-down simulator and a regular STR fed-batch cultivation were compared. In this article we show that the cultivation of Escherichia coli K-12 substrain W3110 in a scale-down two-compartment bioreactor has consequences on the free amino acid pools. Repeated occurrence of high glucose zones affects the biosynthesis of amino acids derived from pyruvate. This is interesting since the pyruvate based amino acids are precursors of the branched chain amino acid pathway, which is further disturbed by an unbalanced regulation caused by a frameshift mutation in the ilvG gene of E. coli K-12 strains. As a consequence the formation of non-typical amino acid norvaline was detected in the two compartment bioreactor cultivation.