Isolation,Identification and Physicochemical Properties of a Lectin from Flos sophorae immaturus
Flos sophorae immaturus lectin(FSIL)was separated by ammonium sulfate precipitation,dialysis,DEAE(Diethylaminoethyl)anion exchange chromatography and Superdex G-75 gel column,and its lectin activity,glycosidic bond type,glycobinding specificity,thermal stability and in vitro digestion characteristics were further studied.The results showed that the relative molecular weight of FSIL was 62.3 ku.FSIL was a glycoprotein by Periodate acid-Schiff staining experiment.The glycosidic bond of FSIL was identified as O-type glycosidic bond by PNGase F enzymatic glycolysis method and reductiveβ-elimination method.FSIL showed coagulation activity for A,B,and O blood types.It was identified as a galectin-type lectin.After heat treat in a boiling water for 30 min,FSIL exhibited non-coagulation activity.Meanwhile,its secondary structure became looser and hydrophobic amino acid residues exposed by circular dichroism and ultraviolet absorption spectra.FSIL was stable in simulated gastric fluid(SGF)and no obvious degradation was observed after digestion for 60 min.FSIL was reasonably stable in simulated intestinal fluid(SIF)with a total digestion time of 40 min.However,after 30 minutes of boiling water treatment,FSIL's resistance to protease digestion was significantly reduced,and it was completely degraded by SGF and SIF within 10 minutes.This study provides a theoretical foundation for the safe development of Flos sophorae immaturus products and the application of FSIL.
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