OPTIMIZATION OF PROTEASE PRODUCTION IN ASPERGILLUS FLAVUS USING TAGUCHI APPROACH AND NANO SUPPLEMENTATION

ABSTRACT Protcases are hydrolytic biocatalysts present in all forms of life which are involved in several biological processes.Microbial proteases are preferred for industrial application due to the rapid growth and ease of genetic modification.However,one of the biggest limitation to its application is its high cost of production.Thus,this research focuses on production of microbial proteases using low cost readily available agro-wastes as substrates. Melon shell,groundnut shell,and groundnut peel were screened for the production of proteases by Aspergillus flavus,Aspergillus terrus,Geotrichumcandidum and an unidentified isolaieunder Submerged method Fermentation(Smf).All the agro-wastes produced with proteases with varying activities;however,Aspergillus flavus gave maximum protease activity of 121.0 U/ml in Groundnut shell under Submerged method Fermentation(SmF).Agricultural wastes (melon shell,groundnut shell,and groundnut peel)used in this study produced proteases hence;they can be used as substitutes to the synthetic substrate which are expensive. The cultural and nutritional conditions for protease production by Aspergillus flavus were optimized using the Taguchi technique and Nano-supplementation with caleium oxide nanoparticles.The Protease produced by Aspergillus flavuswas improved to 145.75U/mL using Taguchi method of optimization and nanoparticleat inoculum size of 0.5ml,fermentation time of 72h,substrate size of 0.5g.pH 12.CaONPs concentration of 0.5ml,and ANOVA showed that the nanoparticle contributed 10.63586 %to improved production. Agricultural waste(Melon Shell,Groundnut Shell,Groundnut Peel)used in this study produced protease hence,they can be used as substitutes to the synthetic substrates which are expensive.This study can also contribute to the attainment of SDGs6(water and sanitation for all)and SDGs12(sustainable consumption and production pattern)