CAMPERI Silvia Andrea
congresos y reuniones científicas
Trypsin partition behavior in poly(ethyleneglycol) and magnesium sulfate aqueous two-phase system
MARIELA M. MARANI; SILVIA A. CAMPERI; OSVALDO CASCONE
Congreso; XXXXIX Reunión Annual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2003
Aqueous Two-Phase Systems (Atpss) is an integrative purification process that combines clarification and coarse purification in one step reducing the downstream processing cost. This extraction technology offers high capacity, high activity yields and easy scale up. Polymer-salt systems have the advantage of low viscosity and low cost compares to polymer-polymer systems.Trypsin is a proteolytic enzyme usually employed in pharmaceutical, food and leather industries as well as in biochemical research. Trypsin behaviour in poly(ethylene glycol) (PEG) and magnesium sulphate aqueous two-phase systems was characterized in order to find suitable systems for its purification.Phase diagrams of PEG-magnesium sulphate for different PEG MW were described. The influence of PEG MW, pH and NaCl on the partition coefficient of trypsin (KTRY) and total proteins (KTP) of a pancreatic extract was studied. In this systems studied, trypsin partition was only slightly influenced by tie-line length and did not show regular trend with pH. The KTRY decreased from 0.4 to 0.15 and de KTP from 0.85 to 0.047 by increasing the PEG MW from 4000 to 10000. The NaCl addition changed dramatically the KTRY from0.15 to 23 but no effect on KTP was evidenced. These results may be considered as an interesting alternative for the use in trypsin purification from crude extract.