Production, Partial Purification and Optimization of Oilseed-Based Protease and its Application as an Efficient Eco-Friendly Alternative for Destaining and Dehairing Process

Proteases are a group of industrially valuable enzymes with significant demand owing to their diverse and extensive range of biotechnological applications. This study aimed to purify and optimize protease from oil seeds as oil seeds contain high oil and protein content along with the enzymes responsible for the digestion of these biomolecules. Three indigenous oil seeds including Brassica compestris, Zea mays and Helianthus annus were included in this study and proteases were isolated and characterized by germinating these seeds at different time periods. The partial purification of enzyme was carried out up to 4.8, 2.5 and 2.44-fold from 5-days old germinating oilseeds of MH5, BH5 and SH5. Total protein content (TPC) of 30, 50 and 250 mgmL-1 were obtained for MH5, BH5 and SH5, respectively with maximum precipitation at 60-70% saturation with (NH4)2SO4. The enzyme was observed to be stable up to 50 oC at pH 8.0 using casein as substrate. The enzyme activity was alleviated using Ca2+, Fe2+, Cu2+ and Co2+ except for BH5 where Ca2+ was ineffective in enhancing or reducing enzyme-activity. The specific-activity values of 0.07, 0.094 and 0.02 mgmL-1 were observed for MH5, BH5 and SH5 respectively. The supplementation of enzyme preparation in detergent completely removed the blood stain from a stained cloth providing the basis of enzyme utilization in detergent industry. Moreover, dehairing effects were observed when applied to goat ear expanding the scope of purified enzyme to enhance the quality of leather in tannery industries by promoting the dehairing process of animal skins thereby avoiding environmental pollution.