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Purification and Characterization of Three Lectins Extracted from Cassia Fistula Seeds and Effect of Various Physical and Chemical Agents on Their Stability
Author(s) -
Ali M. Abbas,
Sayeed M. Abu,
Absar Nurul
Publication year - 2004
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.200400097
Subject(s) - chemistry , sephadex , size exclusion chromatography , chromatography , galactose , affinity chromatography , biochemistry , ion chromatography , enzyme
Three lectins designated as CSL‐1, CSL‐2 and CSL‐3 were purified from Cassia fistula seeds by gel filtration on Sephadex G‐50 followed by ion‐exchange chromatography on DEAE cellulose and finally by affinity chromatography on Sepharose 4B. The molecular weights of the lectins CSL‐1, CSL‐2 and CSL‐3, determined by gel filtration on Sephadex G‐75 column were 37,000, 42,400 and 46,000, and by SDS gel electrophoresis were 37500, 42000 and 46500, respectively. All three lectins agglutinated rat red blood cells and the agglutination was inhibited specifically by galactose and galactose containing saccharide. The neutral sugar contents of the lectins, CSL‐1, CSL‐2 and CSL‐3 were estimated to be 3.5, 3.1 and 2.0%, respectively. The sugar composition of the lectins was found to be galactose in CSL‐1, galactose and glucose in CSL‐2, and galactose and mannose in CSL‐3. The lectins exhibited maximum hemagglutinating activities around pH 7.2 to 7.5 and at a temperature range of 20° to 35 °C. Biological activities of the lectins were abolished sequentially with the increase in concentration of acetic acid and denaturant solutions such as urea and guanidine‐HCl.