Microbial production of multienzyme preparation from mosambi peel using Trichoderma asperellum

Abstract Fruit and vegetable whole sale markets produces huge quantities of wastes in the forms of discarded fruits and vegetables or their parts, creating unhygienic conditions around the area and resulting into environmental pollution. Many enzymes have good applicability in food industries mainly in the production of clarified fruit juices. However, cost of production of such food enzymes are generally high due to costly raw materials. Utilization of fruit and vegetable wastes as carbon sources for enzyme production through microbial intervention has already been reported by various workers. The present study reports the feasibility of using mosambi (Citrus limetta) peel as substrate for multienzyme preparation including pectinase, cellulase and amylase, using a potential fungal isolate.Conditions viz. temperature, pH, incubation time and nutrient addition were optimized for enzymes production from fresh mosambi peel as substrate using Trichoderma asperellum NG-125 (accession number-MW287256). Maximum activities (U ml-1min-1) of pectinase (595.7±2.47), cellulase (497.3±2.06) and amylase (440.9±1.44) were observed at pH 5.5, incubation temperature of 30°C after 10 days of fermentation. Macro-nutrients addition @ 0.1% ammonium sulphate, 0.01% potassium-di-hydrogen-ortho-phosphate enhanced the production of enzymes. The purified pectinase, cellulase and amylase by SDS-PAGE revealed three bands to molecular mass of 43, 66 and 33kDa, respectively.Out of four natural fibre matrices (bagasse, rice husk, paddy straw and wheat straw) tested for immobilization, maximum enzyme activity retention percent was observed on bagasse matrix (pectinase-56.35%, cellulase-77.68% and amylase-59.54%). Enzymatic juice clarification yield obtained with test enzyme was 75.8% which was as comparable to 80.5% of commercial enzyme.