The purpose of this research was to production and characterization protease from patogenic bacteria E scherchia coli. The optimum pH and temperature of protease fr om Escherichia coli were 7.5 and 40 ºC, respectively. Co 2+ (5 mM) and Ba 2+ (5 mM) increased E. coli protease whereas Na + (1 and 5 mM), K + (1 and 5 mM), Mn 2+ (1 and 5 mM), Co 2+ (1 mM) and Ba 2+ (1 mM) inhibited protease from E. coli inhibited the enzyme. Study on the effect of metals ion and specific inhibitors indicated that p rotease from E.coli was serin metaloenzyme. Molecul er weight of protease by using SDSPAGE and zymogram technique fr om E. coli was 36 kDa.