Taming Environmental Bacteria with Synthetic Biology
Environmental bacteria gained attention as promising chassis for Metabolic Engineering and Synthetic Biology. Their robustness and versatility in regards to their metabolisms holds potential for many applications. This versatility is exploited for degradation and production of numerous chemical, while the robustness makes them ideal for big-scale fermentation. However, the toolbox for engineering and manipulating environmental bacteria is still lacking substantially behind the ones of model organisms. Therefore, we focus on developing new and advance and adapt existing tools. Here, we will show several tools for genome engineering in Pseudomonas putida. Firstly, fast streamlined homologous recombination through synthetic control of plasmid replication. This allows targeted insertions, mutation and deletions. Secondly, multiplex base editing for targeted mutation of single bases. Additionally, we present a toolbox to tailor expression of target genes. Taken together, these approaches allow high-speed engineering of a broad range of bacteria and construction of fine-tuned genetic circuits.
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