Fatty Acid Modification of the Anticancer Peptide LVTX-9 to Enhance Its Cytotoxicity against Malignant Melanoma Cells

Spider venom is a valuable resource for the development of novel anticancer drugs. In this study, we focused on novel linear amphipathic α-helical anticancer peptide LVTX-9, which was derived from the cDNA library of the venom gland of the spider Lycosa vittata. The cytotoxicity of LVTX-9 against murine melanoma cells in the range of 1.56–200 μM was tested and found to be significantly lower than those of most anticancer peptides reported. Its IC50 was determined to be 59.2 ± 19.8 μM in a serum or 76.3 ± 12.7 μM in serum-free medium. Fatty acid modification is a promising strategy for improving peptide performance. Therefore, to enhance the cytotoxic activity of LVTX-9, fatty acid modification of this peptide was performed, and five different carbon chain length lipopeptides named LVTX-9-C12-C20 were produced. Among them, the lipopeptide LVTX-9-C18 showed the highest cytotoxic activity in relation to B16-F10 cells, whether in a serum or serum-free medium. Most importantly, the cytotoxic activity of LVTX-9-C18 was improved by about 12.9 times in a serum medium or 19.3 times in a serum-free medium compared to that of LVTX-9. Subsequently, assays including scanning electron microscopy, trypan blue staining, lactate dehydrogenase leakage assay, and hemolytic activity could indicate that the potential direct cell membrane disruption is the main mechanism of LVTX-9-C18 to induce cancer cell death. Furthermore, the LVTX-9-C18 also showed strong cytotoxicity in relation to 3D B16-F10 spheroids, which indicates it might be a promising lead for developing anticancer drugs.

Effects on Physicochemical Characteristics of Yoghurt and Ice Cream with Fatty Acid Modification and Cholesterol Removal

This study investigated the effect of fatty acid modification and cholesterol removal on physico-chemical characteristics of yoghurt and ice cream. Fatty acid profile of milk fat was modified by feeding calcium salts of soybean oil fatty acids to cows and cholesterol was removed by b-cyclodextrin. b-cyclodextrin removed 76% and 60% cholesterol from yoghurt and ice cream. Modification of fatty acid composition did not have a significant effect on a-tocopherol content; while b-cyclodextrin treated milk had substantially lower a-tocopherol content. The concentration of a-tocopherol in control and b-cyclodextrin treated yoghurt was 45.62, 32.73 mg/g and 210.34, 185.56 mg/g for ice cream, respectively. Fatty acid modification and cholesterol removal significantly decreased the overrun in ice cream (P<0.05), with no effect on sensory characteristics of yoghurt and ice cream. These results evidenced that milk with higher content of unsaturated fatty acids and low cholesterol can be used in the formulation of yoghurt and ice cream with improved health benefits and suitable sensory features.