The aim of the current study is to evaluate the effects of medium compositions on the production of protease inhibitor in Streptomyces sp. SMF13. The production of protease inhibitor was counter-currently linked to extra-cellular protease, which were regulated by the culture conditions. Nitrogen source was the most critical ingredient affecting the production of protease inhibitor and protease. Carbon source was an important factor to determine the culture pH which affected very clearly the formation of protease and protease inhibitor. Inorganic phosphate inhibited the protease inhibitor production which was linked to the cell growth rate, although the optimal conditions for the production of protease inhibitor were not favouring to the cell growth.