ISOLATION AND CHARACTERIZATION OF LECTIN FROM SEEDS OF ZIZYPHUS MAURITIANA
Ashwin B. Butle and Mandakini B. Patil*
ABSTRACT
The present paper deals with isolation and characterization of a novel
galactose binding lectin from the seed cotyledons of Zizyphus
mauritiana. The extracted lectin in PBS at pH 7.00 was fractionated by
Ammonium sulphate precipitation. The fraction obtained between 0 –
35% ammonium sulphate precipitation was further purified by affinity
chromatography on guar-gum followed by ion exchange
chromatography on DEAE-cellulose. The isolated lectin was found to
be tripeptide, molecular weight of three different subunits of 38 kD, 25
kD and 19.95 kD on SDS - PAGE. Isolated lectin agglutinated rabbit
erythrocytes showing maximum agglutination with human B+ erythrocytes. It was active
between pH 4.0 - 9.0 with an optimum activity at pH 7.0 and was found fairly stable from pH
4.5 to 8.0. The lectin optimally active at 37°C was stable up to 80°C. Presence of metal ions
such as Mg++ and Mn++ positively influenced the activity of lectin but the activity was
inhibited in the presence of Cd++, Hg++ and Pb++. The lectin showed maximum enzymatic
activity towards the substrate β – oNPGal.
Keywords: Lectin, Zizyphus, erythrocyts, affinity chromatography, agglutinin.
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