Pernisine in an extracellular protease, isolated from archaeon Aeropyrum pernix, which belongs to the group of subtilisin-like proteases. Native persine has the ability to degrade prions which cause Creutzfeldt – Jakob disease. The production of native pernisine from the archea is not economical, mainly because of low yields and demanding growth conditions, which can not be ensured by the regular indutrial scale fermenters. As a part of this research project, we used bacterium Streptomyces rimosus as a host for heterologous expression of the recombinant pernisine. S. rimosus ∆OTC was transformed with a replicative shuttle E. coli – Streptomyces plasmid construct pVF, which contained nucleotide sequence encoding pernisine. The aim of the Master's thesis was to successfully transform the plasmid for the production of recombinant pernisine into S. rimosus. The successful production of pernisine was confirmed by partial isolation of pernisine with affinity chromatography and further analysis with SDS – PAGE. Proteolytic activity of recombinant pernisine was measured with the use of asocasein assay and zymography. We have confirmed successful production of pernisine by the selected transformants of S. rimosus. We have thus confirmed that S. rimosus is suitable host for production of recombinant pernisine. Further evaluation of the properties of pernisine produced this way will be necessary to establish the capability of the recombinant pernisine to degrade infectious prion proteins.
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