One-Step Purification of Melittin Derived from Apis mellifera Bee Venom

The concern over the use of melittin in honey bee venom due to its adverse reaction caused by allergens such as phospholipase A2 (PLA(2)) and hyaluronidase (HYA) has been an obstacle towards its usage. We developed a novel single-step method for melittin purification and the removal of PLA(2) and HYA. This study explores the influence of pH, buffer compositions, salt concentration, and types of cation-exchange chromatography resins on the recovery of melittin and the removal of both HYA and PLA(2). Melittin was readily purified with a strong cation-exchange resin at pH 6.0 with sodium phosphate buffer. It resulted in a recovery yield of melittin up to 93% (5.87 mg from a total of 6.32 mg of initial melittin in crude bee venom), which is higher than any previously reported studies on melittin purification. PLA(2) (99%) and HYA (96%) were also successfully removed. Our study generates a single-step purification method for melittin with a high removal rate of PLA(2) and HYA, enabling melittin to be fully utilized for its therapeutic purposes.