Secretion of biologically active porcine prophospholipase A2 by Saccharomyces cerevisiae

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Abstract
The cDNA coding for pocine pancreatic prophospholipase A2 (proPLA) has been cloned and expressed in Saccharomyces cerevisiae. Expression and secretion of proPLA could only be obtained after fusing the proPLA to the prepro sequence of the yeast α‐mating factor. Upon secretion, the fusion protein was cleaved by the KEX2 protease yielding a 140‐amino‐acid zymogen‐like form of the phospholipase A2. This protein was purified in high yield by ion‐exchange chromatography. Limited proteolysis with trypsin cleaved the ‘zymogen’ to yield active phospholipase A2, which was indistinguishable from the authentic porcine pancreatic enzyme. These results show that a protein with a disulphide bridge content as high as 7 per 124 amino acid residues can be correctly processed by the yeast secretory apparatus.