Extraction and Purification of Protease from Nigella sativa for Its Potential Use in Celiac Disease

Abstract

Celiac disease (CD) is a disease of the digestive system resulting from the intolerance of autoimmune system against gluten and gliadin proteins in wheat, barley, rye and some varieties of oats. In this work protease was isolated and partially purified from Nigella Sativa using ammonium sulphate, acetone and trichloroacetic acid. The purified protease was evaluated further for its possible use in celiac disease by its ability to hydrolyse both, gluten and gliadin. Data indicate that the highest purity of digestive protease enzymes was obtained by acetone 80% and 0.2M trichloroacetic acid (TCA). The enzyme purified by acetone exhibited the high value of Vmax (166.66 mg/min) and the low value of Km (0.133 mg/ml). When the obtained enzyme was tested to hydrolyse gluten and gliadin at two increasing concentrations of 1 and 2 mg/ml, almost similar efficacy was observed (28.46% and 32.02%; for gluten) and (22.7% and 16.84%; for gliadin), whereas Digestin, a positive control which contain Papain and Sanzyme 3500 and works by helping in the breakdown of proteins, produced higher efficacy at 2 mg/ml to hydrolyse gluten (38.67%) with 1 mg/ml (25.68%) whereas similar results were observed against gliadin at both concentrations (22.06% and 22.5%). This study show the efficacy of protease obtained from Nigella Sativa as a natural and economical source for the hydrolysis of gluten and gliadin, which is considered the primary reason for celiac disease.