title: 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 77.59 mM NaCl, pH 5.65, AOT 127.44 mM sodium bis (2-ethylhexyl) sulfosuccinate (AOT) for the forward extraction; and 592.97 mM KCl, pH 8.01 for the backward extraction. The yield was 63.21 ± 2.35 mg 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.
authors: School of Food Science and Engineering, Harbin Institute of Technology, Harbin, Heilongjiang 150090, China; Guelph Food Research Center, Agriculture and Agri-Food Canada, Guelph, Ontario N1G 5C9, Canada.
authors: Shudong
authors: S
authors: He
authors: School of Food Science and Engineering, Harbin Institute of Technology, Harbin, Heilongjiang 150090, China; Guelph Food Research Center, Agriculture and Agri-Food Canada, Guelph, Ontario N1G 5C9, Canada. Electronic address: john.shi@agr.gc.ca.
authors: John
authors: J
authors: Shi
authors: School of Food Science and Engineering, Harbin Institute of Technology, Harbin, Heilongjiang 150090, China; Institut des Régions Arides de Médenine, Laboratoire d'Aridoculture et Cultures Oasiennes, 4119, Tunisia.
authors: Elfalleh
authors: E
authors: Walid
authors: School of Food Science and Engineering, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors: Hongwei
authors: H
authors: Zhang
authors: School of Food Science and Engineering, Harbin Institute of Technology, Harbin, Heilongjiang 150090, China. Electronic address: maying@hit.edu.cn.
authors: Ying
authors: Y
authors: Ma
authors: Guelph Food Research Center, Agriculture and Agri-Food Canada, Guelph, Ontario N1G 5C9, Canada.
authors: Sophia Jun
authors: SJ
authors: Xue
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D037102:Lectins / Q000737:chemistry*; D008823:Micelles; D027805:Phaseolus / Q000737:chemistry*
chemical_list: D037102:Lectins; D008823:Micelles
keywords: Lectin; Purification factor; Response surface methodology; Reverse micellar extraction; Yield