My Homemade Italian Bee Bee Venom Peptide Purification, Structure Identification, Activity Determination, And Solid Phase Synthesis

The earliest recorded investigation of the venom from the honeybee Apis mellifera was in1837. Science then, The European honey bee, Apis mellifera, is one of the most studied insects. As a consequence, its venom is probably the best characterized venom in the Apidae family.Venom produced from the honeybee Apis mellifera is a highly complex mixture which consists of the enzymes phospholipase A2and hyaluronidase, and the peptides melittin, apamine, MCD-peptide, secapin and tertiapin. Bee venom showed many pharmacological activities, such as anti-inflammatory, anti-radiation, anti-bacterial, antihypertensive, analgesic, and anti-viral activitie. Because of its low molecular weight, high stability in denaturing conditions and high spectific activity on the kinds of receptors, they also have been used in studies of structure-function relationship and neurochemical mechanism.In China, Bee venom constituents had become a commercial drug to treat Rheumatoid Arthritis (RA) and Peripheral Neuritisy. To investgate the composition of the bee venom produced in China and discover the new active peptide led us to isolat and identify the chemical constituents of bee venom.In this work, we report the studies on establishing the LC-LTQ-FT MS spectrum of the crude bee venom and the purification and characterization of14peptides, ten of which were purified using kinds of chromatography techniques。while the others were identified by LC-ESI-Q-TOF.The structure of three new peptides were idenfied based on the spectroscopic and chemical method, and named melittin F1secapin-1and secapin-2respectively.Melittins F2, F3, F4,and melittin-G were different breakdown products of melittin or its precursor, identified by LC-ESI-Q-TOF. The existance of these peptides were not reported in literature before.Established the LC-LTQ-FT spectrum of the crude bee venom by HPLC/FTICR-MS. The result indecated that the molecular weights of these peptides almost distributed between1000Da to3500Da. Only four of them are larger then7000. Nine peptides were idenfied based on molecular weights and further investigations. The inhabition of secapin, Api m6.02and Api m6.03on voltage-dependant potassium channel and sodium channel was studiedA secapin-1derivative was synthesized through solid-phase synthesis and purified using kinds of chromatography techniques.