Production and properties of invertase from a Cladosporium cladosporioides in SmF using pomegranate peel waste as substrate

Abstract

ObjectiveTo use suitable fungal strain for production of invertase and to study the effect of external substances and metal ions that may enhances the production of invertase. MethodsThe culture and nutrient requirements of Cladosporium cladosporioides (C. cladosporioides) for production of invertase in a medium at different pH, temperature, incubation period, carbon source and nitrogen source were quantified in present study. ResultsThe optimum pH, temperature and incubation period for enzyme production were 4, 30 °C and 4th day respectively. Among the carbon source pomegranate peel was recorded to be the best carbon source for enzyme production and yeast extract at 1% was ideal nitrogen source for the invertase production. The enzyme was purified by ammonium sulphate precipitation, dialysis and DEAE-Cellulose column chromatography. A trial for the purification of invertase resulted in an enzyme with specific activity of 197.50 Units/mL with 6.35 folds of purification. The purified invertase had a maximum activity at pH 6 and the Km value for pomegranate was 0.26 mg/mL. Analysis of this enzyme for molecular mass was carried out by SDS-PAGE electrophoresis which revealed one band 61 kDa. ConclusionFrom this study, it can be concluded that the pomegranate peel waste can be more effectively used as a substrate for the production of enzyme under optimized culture conditions.