Production and characterization of a-amylase from mango kernel by Fusarium solani NAIMCC-F-02956 using submerged fermentation.
Devendra Kumar, Yadav K. K., Muthukumar M., Neelima Garg
Author Affiliation: Division of Post Harvest Management, Central Institute for Subtropical Horticulture, Lucknow - 227 107, India.
Journal of Environmental Biology 34 : 1053-1058
Abstract : Microbial production of enzymes using low valued agro industrial wastes is gaining importance globally. Mango is one of the major fruit processed into a variety of products. During processing 40-50% of solid waste is generated in form of peel and stones. After decortications of mango stone, kernel is obtained which is a rich source of starch (upto 60%). It was utilized as a substrate for a-amylase production using Fusarium solani. Maximum ?-amylase production (0.889 U g-1) was recorded using a substrate concentration of 5% (w/v), pH-4 and temperature 30°C on 9th day of incubation. Supplementation of production medium with micronutrients viz., Ca2+, Fe2+ or Mg2+ improved the enzyme production while, Zn2+, B3+ or Mn2+ ions exhibited inhibitory effect. The extracellular protein was precipitated by ammonium sulphate up to 70% saturation, dialyzed and purified (27.84 fold) by gel-exclusion (Sephadex G-75) chromatography. Protein profiling on 12% SDS-PAGE revealed three bands corresponding to 26, 27 and 30 kDa molecular sizes. The optimum amylase activity was achieved at pH 5.0 at 40°C. The Michaelis constant (KM), Vmax and activation energy (-Ea) were found to be 3.7 mg ml-1, 0.24 U mg-1 and 42.39 kJ mole-1, respectively.