A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin.
Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9% NaCl at 4 degrees C. The extract obtained was filtered (0.45 microm membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour.
Fasina
Yewande O
YO
Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon S7N 5A8, Saskatchewan, Canada. fasinyo@acesag.auburn.edu
Swaisgood
Harold E
HE
Garlich
Jim D
JD
Classen
Henry L
HL
D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D010865:Pilot Projects; D037121:Plant Lectins / Q000302:isolation & purification*; D030262:Soybean Proteins / Q000302:isolation & purification*; D013025:Glycine max / Q000737:chemistry; D014462:Ultrafiltration; D010835:Phytohemagglutinins
D037121:Plant Lectins; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins