Rhizosecreted proteases inhibition for the improvement of recombinant protein production in Arabidopsis thaliana

Plant-based biopharmaceuticals have gained a lot of interest in the past decade due to their reduced cost and relative safety compared to mammalian cell cultures. While the first plant-made recombinant proteins are now reaching the market, the production systems still need improvements to maximize their competitiveness. Optimizing production hosts requires the identification and subsequent inhibition of the most active endogenous peptidases, proteolysis being one of the main factors limiting yields.
The aim of our study was to identify root-secreted proteases of Arabidopsis thaliana involved in target protein degradation (BSA) and inhibit them in vivo. Biochemical analyses identified serine proteases as the main class responsible for BSA degradation. An RT-qPCR experiment led to the choice of the serine protease gene SBT4.12 and its homologs as targets for an amiRNA-mediated silencing approach. Arabidopsis amiRNA-expressing lines showed lower levels of expression for SBT4.12 and reduced proteolytic activity in their rhizosecreted extracts. Crossing these lines with recombinant protein producing lines could lead to an improved production platform for proteins of interest.