Purification and partial characterization of a fructose-binding lectin from the leaves of Euphorbia helioscopia

ABSTRACT

A lectin was purified from leaves of Euphorbia helioscopia, by a combination of ion-exchange and gel filtration chromatography. On ion exchange using a DEAE- cellulose column in 0.2 M phosphate buffer, pH 7.2, the bound protein was eluted with a linear sodium chloride gradient of 0.1 M to 0.5 M. Further purification of the lectin was achieved by gel filtration on Sephadex G-100. Euphorbia helioscopia lectin [EHL] agglutinates only chick erythrocytes, showing no agglutination of all human blood group erythrocytes. The EHL induced hemagglutination is inhibited by fructose. The purified protein showed one band, both in non-denaturing PAGE and SDS-PAGE establishing the charge and size homogeneities of the lectin preparation. The molecular mass of the lectin as indicated by SDS-PAGE was approximately 31 kDa and that estimated from G-100 gel filtration chromatography was about 65 kDa establishing that the lectin is a homodimer. The lectin was stable within a temperature range of 0°C-40°C and exhibited a narrow range of pH stability, being optimally active at around pH 7. EHL also possesses antimicrobial activity and is an inhibitor of bacterial growth particularly Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli