Acid phosphatase is one of the most important enzymes in venom of honeybees that has many medical and pharmaceutical applications. In this study, an acid phosphatase was isolated and homogeneously purified from venom of Apis mellifera lamarckii that was named BVACP. The purification process was carried out using column chromatography on CM-cellulose and Sephacryl S-300 resins. BVACP specific activity of 305.4 Umg^-1 was obtained with 33.2% recovery and 3.9 times purification was realized. BVACP was found monomer protein of 46 kDa by gel electrophoresis and gel filtration chromatography. BVACP displayed its maximum activity at pH 5.0, with K_m value of 3.3 mM p-NPP and Vmax was 0.45 Umg^-1. It was activated with MnCl₂, MgCl₂, CoCl₂, NiCl₂ and CaCl₂, while FeCl₂, CuCl₂ and ZnCl₂ inhibited it. Ammonium molybdate and sodium fluoride inhibited BVACP non-competitively with Ki values of 1.2 µm and 0.37 mM, respectively. BVACP had diverse inhibition ratios against specific bacterial strains and had no effect on others, referring to a selective spectrum of activity. It selectively and potently affects A549, HOS and HCT116 tumour cells. These potent antitumor and antibacterial activities of BVACP could be useful for many clinical and medical applications.