Investigation of enzymatic activities of Apis mellifera species bee venom

Our country, with its ecological richness, hosts 5 pure honey bee races (Anatolian bee, Caucasian bee, Carniolan bee, Iranian bee and Syrian bee) and many different ecotypes (Gökçeada bee, Muğla bee, Yığılca bee, Efe bee etc.). Honey bee races and ecotypes have different physiological, morphological and behavioral characteristics. Bee venom is a complex mixture of enzymes, low-molecular compounds and peptides produced by honey bees from their venom glands. It contains phospholipase A2, phospholipase B, hyaluronidase, phosphatase and α-glucosidase peptides. It also contains bioactive components such as melittin, apamin, mast cell degranulating peptide, adolapin, tertiapin, secapin and cardiopep. In this study, the raw of Apis mellifera L. was collected from three different regions (Ankara, İzmir, and Mersin) using the electrical stimulation method. The components were separated using chromatographic and electrophoretic methods, and the peptide profiles of the mixtures were analyzed. The proteolytic activity of the raw was determined using zymogram analysis with gelatin and casein substrates. Furthermore, the hyaluronidase and phospholipase activities of the raw were investigated. According to HPLC results of Ankara, İzmir and Mersin venoms, the fraction numbers are at least 30, 52 and 40, respectively. In SDS-PAGE analysis, all three localities showed similar protein bands. In zymogram analysis, caseinolytic activity was observed in the range of 37-52 kDa at pH 7.4, and in addition, caseinolytic activity was detected in the range of 30-37 kDa in İzmir venom. Gelatinolytic effect was determined only in İzmir venom at around 95-130 kDa and 270 kDa. In the phospholipase test, PLA2 activity with a zone diameter of 1 cm and hyaluronidase activity in the range of 6.5-16 kDa were observed in all three venoms after 16 hours of incubation. The findings of this study highlight the proteolytic, lipolytic, and hyaluronidaseactive peptides present in honeybee venom, demonstrating its potential for medical and commercial applications. The venom, even within the same bee species across different localities, shows potential for structural variations due to intraspecific differences. These enzymatically active components may serve as valuable tools for understanding activation mechanisms