Studies On Lens Culinaris B Galactosidase Purification Immobilization And Its Applications

Abstract

and#61656; and#946;-Galactosidase from Lens culinaris (Lsbgal) seeds were purified to apparent electrophoretic homogeneity. The enzyme was purified to 857 fold with a specific activity of 87 U/mg. newlineand#61656; Mr of Lsbgal under reducing and denaturing condition as determined from SDS-PAGE showed heterodimeric bands of 45 and 30 kDa, respectively. newlineand#61656; The Mr of native Lsbgal was calculated from Size-Exclusion Chromatography and was found to be 76 kDa. newlineand#61656; The optimum pH and temperature were found to be 3.0 and 58 and#8451;, respectively. newlineand#61656; ONPG was hydrolyzed at highest rate and Lsbgal showed a Km of 1.21 mM with it. newlineand#61656; Purified Lsbgal showed successful synthesis of Galacto-oligosaccharides (GOS) via transgalactosylation reaction when incubated with higher amount of lactose (200 g/L). newlineand#61656; Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC) were carried out to analyze the products, which showed trisaccharides as a major products among total GOS. newline