Staphylokinase (SAK) is abacterial kinase produced by certain strains of Staphylococcus spp. It is a 14.5 kDa, consisting of 136 amino acids single polypeptidechain protein, which activates plasminogen to form plasmin and digest fibrin clots. SAK is a promising thrombolytic agent for the treatment of myocardial infarction. The present research was carried out to produce SAK from locally isolated lysogenic Staphylococcus aureus. The main focus of this work was to isolate and characterize lysogenic S. aureus from various environmental samples involving non-pathogenic Staphylococcus spp., while the earlier isolation was done using pathogenic samples.