Elimination of ribosome inactivating factors improves the efficiency ofBacillus subtilis and Saccharomyces cerevisiaecell-free translational systems

Cell-free translational systems based on cellular lysates optimized for in vitro protein synthesis have multiple applications both in basic and applied science, ranging from studies of translation regulation to cell-free production of proteins and ribosome-nascent chain complexes. In order to achieve both high activity and reproducibility in a translational system, it is essential that the ribosomes in the cellular lysate are enzymatically active. Here we demonstrate that genomic disruption of genes encoding ribosome inactivating factors – HPF in Bacillus subtilis and Stm1 in Saccharomyces cerevisiae – robustly improve the activities of bacterial and yeast translational systems. A possible next step in developing strains for production of even more efficient cell-free translation systems could be achieved by combining a genomic disruption of the ribosome hibernation machinery with inactivation of the genes responsible for proteolysis and RNA degradation.