Study On Separation And Purification Of Peptide From Rape Bee Pollen And Its Immunocompetence

There are many kinds of proteins and peptides in the bee pollen.Lots of them have high nutritional value and perfect health function.Recent studies have demonstrated their multiple pharmacological actionssuch as immunomodulatory effects, anti-cancer effects, protection of liverand so on. So, studies of proteins and peptides in the bee pollen are veryvaluable.A peptide named BPP from rape bee pollen proteins which weresalted out by ammonium sulfate (the concentration was 60-80ï¼…) had beepurified. The purification procedures included Superose 12 chromato-graphy, DEAE Sepharose FF ion-exchange chromatography, Q sepharoseF.F. ion-exchange chromatography and Sephacryl S-100 chromatography.Its physicochemical properties and pharmacological effects werestudied. The purity of the bioactive peptide was demonstrated by HPLC.SDS-PAGE analysis showed that the peptide had a molecular weight of 36.3kDa. The isoelectric point of the peptide was estimated to be 6.0 byisoelectric focusing electrophoresis. The wavelength of maximum absorp-tion at approximately 278nm can be detected. Phenol-Sulphuric acidmethod had been used for measuring the total carbohydrate of the peptideand the result showed that the peptide was not glycoprotein.In this study, we investigated the effects of BPP on the immunefunctions of lymphocytes. Proliferation of cells in response to BPP wasmeasured using colorimetric MTT assay. Results showed that BPP(0.0625-2mg/mL) could significantly induce lymphocytes proliferation.And BPP at concentration of 0.25mg/mL had the best effect. From theresults, it was concluded that BPP possesses high immunologicalactivities.