Isolation and Antibacterial Activity of Honey Bee Venom Bioactive from Apis cerana

Abstract

Honey bee venom from Apis species has garnered considerable attention in medicine due to its bioactive, which have antimicrobial activity. This research aims to isolate bioactive melittin and phospholipase A2 (PLA2) of honey bee venom from Indonesian Apis cerana and examine the antibacterial activity of these bioactive. Honey bee venoms were isolated from four locations using an electric shock method and purified using Fast Protein Liquid Chromatography. Purified protein was analyzed by SDS PAGE and measured by the Lowry method. Further, the antibacterial activity of melittin and PLA2 against Gram-positive (Staphylococcus aureus and Salmonella typhi) and Gramnegative (Escherichia coli) bacteria was defined by inhibition zone. Our results show that the average of all bee venoms obtained from four locations with eight sampling times was 17.1 mg proteins. Melittin was purified using gradient steps with 55%, 90%, and 100% elution buffer, while PLA2 was purified using 55% elution buffer. Melittin and PLA2 showed molecular weights of less than 7.5 kDa and 19 kDa, respectively. Protein concentrations in melittin and PLA2 were 95 µg/mL and 142 µg/mL, respectively. Among the two bioactive, only PLA2 showed antibacterial activity. The diameter of the inhibition zone was 7.76 mm when the concentration of PLA2 was 45 µg/mL against E.coli. This result showed that PLA2 from Indonesian Apis cerana has antibacterial activity and may be a promising candidate for the antibacterial agent on Gram-negative bacteria. These findings may contribute to developing new antibacterial substances derived from Indonesian Apis cerana.