Resumen de Engineering mycoplasma species for biotechnological and biomedical applications
Mycoplasmas are a group of bacteria characterized by minimized genomes, limited biosynthetic capacities, and simplified metabolic networks. In this work, we explored how Mycoplasma species can be exploited for biotechnological and therapeutic applications. In chapter two, we engineered M. pneumoniae transposons so that they could efficiently transform different Mycoplasma species. This fact allows the generation of essentiality studies in these species, a critical analysis to identify genes that could be deleted to create attenuated vaccination strains. Chapter three developed a new method that uses high-resolution transposon and proteomics data to infer active metabolic pathways within a cell at a specific moment. This information can be used when engineering attenuated strains. In chapter four, we explored the capacity of M. pneumoniae to express functional human biologics in vitro and in mice lungs. Chapter five used the protein design algorithm FoldX and ModelX to mutate a human interleukin to enhance its properties in terms of affinity to its receptors and increased bacterial expression. Chapter six identified the secretion signals and designed synthetic promoters in the recent-discovered fast-growing Mycoplasma feriruminatoris. Altogether this thesis develops the tools for exploiting Mycoplasmas for biotechnological purposes (M. agalactiae, M. feriruminatoris) and validates M. pneumoniae for human lung therapy immunomodulation.
