Optimized extraction of a lectin from Crataeva tapia bark using AOT in isooctane reversed micelles

Abstract

Crataeva tapia bark lectin was extracted from a crude extract into a reversed micelle phase of the anionic surfactant AOT in isooctane and back-extracted, to a final aqueous phase by addition of butanol. The effects of pH, ionic strength and surfactant concentration on the protein transfer process from the aqueous to the organic phase were characterized, being the best results obtained after 5 min of contact, under agitation, between the two phases, at pH 5.5 (10 mM citrate-phosphate buffer), 30 mM NaCl, and 5 mM AOT. Recovery to a new aqueous phase was performed with 5 min of contact, under agitation, 10 mM citrate-phosphate buffer at pH 5.5, 500 mM KCl and 5% of butanol. The overall yield obtained for the process was 80% for lectin activity and 56% for protein recovery. The efficiency of the process was confirmed by SDS-PAGE analysis.