Purification and charcaterization of extracellular cysteine protease Article

Purification and charcaterization of extracellular cysteine protease inhibitor, ECPI2, from chlorella - Article Example First, they cultured a strain of Chlorella sp. 4533, separated the filtrate by centrifuge, and concentrated it through evaporation. After assaying the protease activity, they eluted the inhibitor and obtained two active fractions, one of which was the primary research component, ECPI-2. The active fractions of this were pooled, dialyzed and concentrated, and then the protein concentration and carbohydrate content were determined and measured. 4. Discussion of Figures and Tables. Table 1 is the purification summary for ECPI-2, providing comparison of progressively purified elements in terms of protein concentration, total and specific activity, and percentage yield from crude of each step in the purification process. This was performed to purify the inhibitor and demonstrate the increasing level of activity. The first step used a DEAE-cellulose column of 3.5x60 cm and quadrupled the specific activity. Next, after the active elements were pooled, dialyzed and concentrated, they were applied to a Sephadex S-300 column (2x130 cm) which increased specific activity by a factor of almost 5X. Finally, after another evaporator concentration, the inhibitor was applied to a 1x150 cm column of Butyl Toyopearl 650 M, again doubling the specific activity; from crude to final, activity was increased by over 40X, giving the authors evidence of purity.