Expression, purification and biophysical characterization of recombinant phospholipase PLA2 overproduced in .

The main purpose of this work was to develop new protocols for high yield purification of secretory phospholipase A2 (PLA2) and to investigate its biophysical properties. We have used a expression system for PLA2 expression and two-stage chromatography for its purification. The biophysical properties of PLA2 were investigated by circular dichroism. A scalable method for high yield purification of recombinant PLA2 was developed. The PLA2 from was expressed in the methylotrophic yeast . Functional active phospholipase A2 with specific activity 73 U/mg was purified with a concentration of at least 3 mg/mL. The role of different divalent ions in PLA2 thermostability were evaluated. Ca and Ba ions significantly increased thermostability of the enzyme.