EXPRESSION AND PURIFICATION OF RECOMBINANT LYSOSTAPHIN IN ESCHERICHIA-COLI

A 1.5 kb plasmid-encoded lysostaphin gene fragment of Staphylococcus s taphylolyticus was amplified by polymerase chain reaction (PCR) and cl oned in Escherichia call by using plasmid pET29b(+) as an expression v ector. By optimizing culture conditions, the activities of lysostaphin were expressed as 66%, 30%, and 4% in extracellular, intracellular, a nd periplasmic fractions of recombinant E. coli, respectively. The enz yme was purified to homogeneity by using a simple one-step fractionati on on bacterial cells of lysostaphin-resistant Staphylococcus aureus m utant. The recombinant enzyme had an Mr of approximate 27 kDa, and its bacteriolytic activity was indistinguishable to the authentic lysosta phin purified from Staphylococcus staphylolyticus.