The present study focuses on the production ofstaphylokinase (SaK) from Staphylococcus hominis MSD1 which is isolated from a new source, kadi (a traditional food). Out of 12 isolates from fermented sample, single potent strain was found to be an efficient producer of SaK which was screened by heated plasma and skim milk agar assay. The purified enzyme showed a maximum specific activity of 2994 U mg-1 and zone of hydrolysis on radial caseinolytic and heated plasma assay.The blood clot lysis activity was compared with the standard, concluded that minimum 0.18 U (10μL) of purified SaK was required to dissolve the blood clot. The purity and molecular weight of SaK was analysed by SDS-PAGE and found to be approximately around 15 KDa.The present study is the first to report SaK from Staphylococcus hominisisolated from kadi, an traditional food. © 2015, Sphinx Knowledge House. All right reserved.