| Summary: | In this work, we pursued the biological characterization of the venom of <i>Montivipera bornmuelleri</i>, a viper from the Lebanese mountains. In relation to its antibacterial potential, the inhibitory effect of this venom on the F<sub>1</sub>F<sub>0</sub>-ATPase enzymes of Gram-positive <i>Staphylocoocus epidermidis</i> and Gram-negative <i>Escherichia coli</i> bacteria was examined. In order to determine the degree of cytotoxicity of the venom on the HCT116 human colon cancer cell lines, the biological MTT proliferation and cell viability test were implemented. After validation of the enzymatic F<sub>1</sub>F<sub>0</sub>-ATPase model by the spectrophotometric method, using quercetin as the reference ligand, results revealed that <i>M. bornmuelleri</i> venom is able to inhibit the activity of the enzyme of these two bacteria with a concentration of the order of 100–150 µg/mL. In addition, a venom concentration of 10 µg/mL was sufficient to kill the totality of HCT116 cell lines cultivated in vitro. These data show that <i>M. bornmuelleri</i> venom is a mixture of diverse molecules presenting activities of interest, and is a potential source to explore in order to discover new drug candidates.
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