ISOLATION AND PHYSICOCHEMICAL CHARACTERIZATION OF A PHYTOLECTIN FROM PISLIM SATIVIIM L. VAR SIKHAR.B. LAI. AND K.D. SAXENA
A phytolectin was isolated from the seeds of Pisum sativum by ammonium sulphate precipitation at 70% saturation which are dialyzed and purified against distilled water. The heterogeneity of lectin studied by polyacrylamide gel electrophoresis at pH 8.2 and 9.2 and was separated in two fractions. The PSL strongly agglutinate blood group A & B, weakly agglutinate AB and not with 0 blood group. Hem- agglutination activity was inhibited by mannose and glucose even in very low concentration, indicating that they possess at least two binding sites. The solubility of PSL revealed that the fraction was soluble in presence of 0.6 M perchloric acid, Na2HPO4 but insoluble in 20% trichloroacetic acid, 5% phosphotungestic acid, saturated NaH2PO4solution. The lectin gave positive molish test, Bial's orcinol, xanthophoretic, biuret and millions reaction. PSL does not detectable amount of sulphur amino acids but are rich in aspartic acid and threonine. The quantitative carbohydrate analysis showed, PSL contained less than 0.5 % carbohydrate and chromatographic analysis reveals that glucose is the main sugar and xylose in traces. Mn and Ca were found to 0.12 % and 0.19 % respectively and trace amount of iron. The agglutination activity depends on the pH, more acidic or basic pH decreases the activity and stability, it was maximum between pH 6.2 to 9.1 for PSL. The hem-agglutination activity of PSL is also dependent to the temperature and it was maximum between 20°C to 30°C and almost zero at 70°C and above temperature.
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