Transcriptional profiling of the stringent response mutant strain E. coli SR reveals enhanced robustness to large‐scale conditions

Large‐scale processes suffer from process performance loss. E. coli SR exhibits a reduced short‐term transcriptional response when exposed to repeated nitrogen starvation in a scale‐down reactor. Summary In large‐scale fed‐batch production processes, microbes are exposed to heterogeneous substrate availability caused by long mixing times. Escherichia coli, the most common industrial host for recombinant protein production, reacts by recurring accumulation of the alarmone ppGpp and energetically wasteful transcriptional strategies. Here, we compare the regulatory responses of the stringent response mutant strain E. coli SR and its parent strain E. coli MG1655 to repeated nutrient starvation in a two‐compartment scale‐down reactor. Our data show that E. coli SR can withstand these stress conditions without a ppGpp‐mediated stress response maintaining fully functional ammonium uptake and biomass formation. Furthermore, E. coli SR exhibited a substantially reduced short‐term transcriptional response compared to E. coli MG1655 (less than half as many differentially expressed genes). E. coli SR proceeded adaptation via more general SOS response pathways by initiating negative regulation of transcription, translation and cell division. Our results show that locally induced stress responses propagating through the bioreactor do not result in cyclical induction and repression of genes in E. coli SR, but in a reduced and coordinated response, which makes it potentially suitable for large‐scale production processes.