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Purification of a lectin from Winged bean ( Psophocarpus tetragonolobus (L.)DC) tubers by affinity chromatography
Author(s) -
Shet Manjunath S.,
Madaiah M.
Publication year - 1987
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.2740410311
Subject(s) - lectin , sephadex , size exclusion chromatography , molecular mass , affinity chromatography , galactose , chemistry , biochemistry , chromatography , sepharose , glycoprotein , enzyme
A lectin from the tuber of winged bean, Psophocarpus tetragonolobus (L.) DC, has been isolated and purified by affinity chromatography using D ‐galactose linked Sepharose‐6B, followed by CM‐Sephadex C‐50. The homogeneity of the lectin was confirmed by PAGE at pH 4.5, SDS—PAGE and gel filtration. It is a glycoprotein with a relative molecular mass (M r ) of 29 000±1000 and 46 000±2000 by SDS—PAGE and gel filtration on Biogel P‐150, respectively. SDS—PAGE in the presence of 2‐mercaptoethanol gave two closely migrated protein bands corresponding to a molecular mass of 14 000 and 15 000. The total carbohydrate content was found to be 7%. The tuber lectin agglutinates both trypsinised and untrypsinised erythrocytes of human blood groups (A, B and A B), rabbit and rat but not sheep and human O group erythrocytes. The lectin is inhibited by D (+) galactose and galactose containing di‐ and trisaccharides. o‐ and p‐Nitrophenyl α‐galactopyranosides are found to be the most potent inhibitory sugars.