Production, purification and characterization of D-aspartate oxidase from the fungus Trichoderma harzianum SKW-36

Abstract

Microbial cellulases are the enzymes widely studied due to their enormous applications in biochemical industry. Among 12 fungal isolates isolated from mangrove plant debris and soil sample collected from Valanthakad Mangroves, Kerala, India, 3 of them were found to exhibit cellulolytic activity. Among them, the most potent isolate which exhibited maximum cellulolytic activity was identified as Trichoderma viride VKF3 [Gene bank accession number- JX683684.1] based on colony morphology, microscopic observation and molecular centeracterization using D1/D2 region amplification. The isolate T. viride VKF3 was found to be non-phytopathogenic against the selected plants. Neighbour joining tree depicted its least divergence rate from the root taxon HM466686.1. T. viride VKF3 was grown under dynamic carbon, nitrogen sources, pH and temperature of the medium to draw out the optimum conditions for cellulase production. Protein stability kinetics and biomass production was also studied upto 11th day of incubation. It was evident from the study, that dextrose and beef extract could be used as major carbon and nitrogen sources in submerged fermentation at pH 9.0 and incubation temperature of 25°C to get maximum CMCase yield. Optimum enzyme recovery period was identified between 5th to 9th days of incubation beyond which the enzyme activity was reduced. By comparing two fermentation methods, submerged fermentation was found to be the best for maximum enzyme production. But utilization of substrates like sugarcane bagasse and cassava starch waste in the SSF offers a better scope in biodegradation of solid waste contributing to solid waste management.