The (phospho) lipase from Staphylococcus hyicus: Expression in Escherichia coli, large-scale purification and application in esterification reactions

The gene coding for the mature part of the (phospho)lipase from Staphylococcus hyicus has been cloned and overexpressed in Escherichia coli. The recombinant enzyme accumulated in the cytoplasm and was purified using a combination of cation- and anion-exchange chromatography and refolding from guanidine/HCl. The purification protocol yielded 0.46 g of pure S. hyicus (phospho)lipase from 100 gram wet E. coli cells. Although the recombinant enzyme has an N-terminal extension of 35 amino acids, the activity towards lipids and phospholipids of different chain-length is identical to that of the wild-type S. hyicus (phospho)lipase. Application of the (phospho)lipase in esterification reactions demonstrated that the enzyme is inactive in microemulsions of charged detergents, but active in the presence of the neutral detergent Triton X-100 and in the absence of a detergent. These results were compared to the performance of a series of other lipases from microbial origin. The results are discussed in view of the possible application of the S. hyicus (phospho)lipase in the modification of phospholipids.