Purification of recombinant phospholipase A₂ enzyme in escherichia coli / Noor Akma Mohamed Sukri

Phospholipase A₂ is a lipolytic enzyme that specifically hydrolyze sn-2 fatty acyl ester bond of phospholipids to yield free fatty acids and lysophospholipids. It was widely use in the several industry including pharmaceutical, food and biotechnology. Therefore, this study was done to purify the phospholipas A₂ enzyme by using hybrid protocol. This method consist of two combination method which is denaturing and native protocol and has ability to purified and retained biological activity of the desired protein. There were two clones involved in the study which is pBADTOPO pla, clone 5 and pBADTOPO pla, clone 8. They were purified at the 37 °C. Result indicated, only small amount of bioactive protein were recovered. Poor recovery of bioactive protein from inclusion bodies may result from the loss of secondary structure during solubilization procedure and interaction among the denatured protein molecules during refolding.