Reverse micellar extraction of lectin from black turtle bean (Phaseolus vulgaris): Optimisation of extraction conditions by response surface methodology

Abstract

Lectin from black turtle bean (Phaseolus vulgaris) was extracted and purified by reverse micellar extraction (RME) method. Response surface methodology (RSM) was used to optimise the processing parameters for both forward and backward extraction. Hemagglutinating activity analysis, SDS–PAGE, RP-HPLC and FTIR techniques were used to characterise the lectin. The optimum extraction conditions were determined as 77.59mM NaCl, pH 5.65, AOT 127.44mM sodium bis (2-ethylhexyl) sulfosuccinate (AOT) for the forward extraction; and 592.97mM KCl, pH 8.01 for the backward extraction. The yield was 63.21±2.35mg protein/g bean meal with a purification factor of 8.81±0.17. The efficiency of RME was confirmed by SDS–PAGE and RP-HPLC, respectively. FTIR analysis indicated there were no significant changes in the secondary protein structure. Comparison with conventional chromatographic method confirmed that the RME method could be used for the purification of lectin from the crude extract.