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bioRxiv · 10.1101/424044

Elimination of ribosome inactivating factors improves the efficiency of Bacillus subtilis and Saccharomyces cerevisiae cell-free translational systems

Brodiazhenko, T.·Johansson, M.·Takada, H.·Nissan, T.·Hauryliuk, V.·Murina, V.
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Abstract

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.

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Brodiazhenko, T., Johansson, M., Takada, H., Nissan, T., Hauryliuk, V., Murina, V.. 2018-09-22. Elimination of ribosome inactivating factors improves the efficiency of Bacillus subtilis and Saccharomyces cerevisiae cell-free translational systems. https://doi.org/10.1101/424044

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