Extraction,Purification and Identification of Lectin from Phaseolus vulgaris

The Phaseolus vulgarisl was used as the raw material to extract lectin,based on the single factor experiment,the extraction process was optimized by response surface experiment. Samples were further purified by ammonium sulfate fractionation,Sephadex G-50 and DEAE-A50 anion exchange chromatography,the purified protein was identified by SDS-PAGE electrophoresis and LC-MS/MS. Results showed that,the optimum extraction conditions were determined as the ratio of material to liquid 1∶13 g/mL,pH7.4 and extraction time 18.5 h the hemagglutinating activity of lectin was(13.26±0.26)×10³ HU/mg.The hemagglutinating activity of the purified target protein reached(59.93±0.31)×10³ HU/mg,the target protein was determined as lectin and the molecular weight was around 31 kDa.