A New Approach to Bioprocess Control by the Analysis of Regulatory Networks (original) (raw)
1997, IFAC Proceedings Volumes
For optimization of recombinant fermentations both the type of expression system and the metabolic capacity of the host cell have to considered. The estimation of the metabolic load on the host cell during expression of recombinant protein is a crucial parameter and gives significant information to approach optimum yield. Metabolic control of microbial cells is based on a hierarchical organisation comprising a multitude of regulatory networks with specific signal molecules linked to specific transduction pathways. Thus the concentration of these specific signal molecules reflects the state of individual regulatory blocks and monitoring of these molecules offers novel tools to optimize control of recombinant ferrnentations. The signal molecule of the stringent control network, guanosinetetraphosphate (ppGpp) was analysed to estimate the metabolic load of different expression systems on the host cell during production of recombinant human superoxidedismutase. The ppGpp concentration was quantitated by ion pair HPLC. Due to the high turnover of ppGpp the development of a special sampling procedure was essential. The formation of ppGpp in relation to various environmental changes and to recombinant protein expression is shown.
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