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Background  Bacteria are widely used as hosts for recombinant protein production due to their rapid growth, simple media requirement and ability to produce high yields of correctly folded proteins. Overproduction of recombinant proteins may impose metabolic burden to host cells, triggering various stress responses, and the ability of the cells to cope with such stresses is an important factor affecting both cell growth and product yield.    Results  Here, we present a versatile plasmid-based reporter system for efficient analysis of metabolic responses associated with availability of cellular resources utilized for recombinant protein production and host capacity to synthesize correctly folded proteins. The reporter plasmid is based on the broad-host range RK2 minimal replicon and harbors the strong and inducible XylS/ Pm  regulator/promoter system, the ppGpp-regulated ribosomal protein promoter P rpsJ , and the σ 32 -dependent synthetic tandem promoter P ibpfxs , each controlling expression of one distinguishable fluorescent protein. We characterized the responsiveness of all three reporters in  Escherichia coli  by quantitative fluorescence measurements in cell cultures cultivated under different growth and stress conditions. We also validated the broad-host range application potential of the reporter plasmid by using  Pseudomonas putida  and  Azotobacter vinelandii  as hosts.    Conclusions  The plasmid-based reporter system can be used for analysis of the total inducible recombinant protein production, the translational capacity measured as transcription level of ribosomal protein genes and the heat shock-like response revealing aberrant protein folding in all studied Gram-negative bacterial strains.    Electronic supplementary material  The online version of this article (10.1186/s12934-019-1128-7) contains supplementary material, which is available to authorized users.

microbial cell factories

Construction and characterization of broad-host-range reporter plasmid suitable for on-line analysis of bacterial host responses related to recombinant protein production