Comparison of extraction methods for isolating kafirin protein from food grade sorghum flour

Abstract

Isolated sorghum (Sorghum bicolor) storage proteins (kafirins) have been successfully used in the production of several bio-materials including adhesives, films, micro-particles, fibers, and biological scaffold material. Comparatively little research has been conducted on the use of isolated kafirins in food products or to produce bioactive peptides via hydrolysis for nutritional uses. To support such research, the aim of this study was to compare existing methods for bulk isolation of sorghum kafirins with the goal of identifying a solvent with the least toxicity that maintained a high extraction rate from food grade sorghum flour. A secondary goal was to characterize the kafirin isolates produced from various extraction methods to provide some information on their potential use in food products to guide future research in this area. Five different extraction methods were compared including 1) aqueous ethanol containing NaOH and sodium metabisulfite, 2) glacial acetic acid, 3) aqueous ethanol with sodium metabisulfite, 4) aqueous ethanol at acidic pH, and 5) alkaline pH alone. The protein contents of the kafirin isolates obtained by the five methods ranged from 49.76% to 56.83%. Kafirin isolates were characterized using reversed phase (RP)-high performance liquid chromatography (HPLC), which revealed substantial variability in the various kafirin patterns among the extraction methods tested. However, characterization of the kafirin isolates by size exclusion chromatography (SEC) did not show a high degree of variability among the methods tested. Likewise, analysis of the samples using sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) showed essentially the same band profiles but with different band intensities among kafirin extraction methods. Surface hydrophobicity of the kafirin isolates varied considerably with isolates extracted with glacial acetic acid and aqueous ethanol plus sodium metabisulfite the most hydrophobic as indicated by hydrophobic dye binding.