The aim of the master's thesis was to improve the efficiency of the isolation of recombinant pernisine from the Streptomyces rimosus expression system by correctly selecting and optimizing the procedures for the isolation and describing the physical properties and enzymatic activity of recombinant pernisine with regard to various parameters such as temperature and pH. Streptomyces rimosus M4081 was transformed to express recombinant pernisine, transformed with plasmid pVF tcp830 srT srT pernisine CO HT. The first step in the isolation of pernisine was performance of Ni2+affinity chromatography, followed by the steps of size exclusion chromatography, dialysis and concentration of the pernisine sample, thereby obtaining even better purity of pernisine, as demonstrated by NaDS-PAGE and zymography. Physical properties were analyzed by CD-spectrometry and fluorescence emission spectrometry, and it has been demonstrated that recombinant pernisine maintains structural conformation well at a temperature range of 25 to 60 °C and in a pH range of 3.0 to 12.0. The enzyme activity of recombinant pernisine was tested with an azocasein test and it has been demonstrated that recombinant pernisne is thermostable because it retains the residual enzymatic activity greater than 90 % at a temperature of 80 °C over a period of 240 minutes. Recombinant pernizine maintains the ramaining enzymatic activity that is greater than 80%, in a wide range of pH from 5.3 to 10.1 at a temperature of 100 °C with the maximum at pH 6.95 and at a temperature of 100 °C. In conclusion, we have shown that the value of the enzymatic activity of recombinant pernisine, at certain levels of isolation, corresponds to the level of purity of the pernisine obtained by the individual levels of isolation.