Bacillus circulans MTCC 7906 aided facile development of bioconjugate nano-silica alkaline protease formulation with superlative dehairing potential

Abstract

The tannery industries utilize environmentally hazardous chemicals to achieve dehairing of animal hides, which causes enormous waterbed pollution & high TDS load. Alkaline protease enzyme for dehairing can be an effective solution to resolve the environmental problems of the tannery industry waste. However, stable, cost-efficient and eco-benign formulations of alkaline protease need to be developed for commercial applications in the tannery industry. This works aimed at development of a nano-formulation of the enzyme alkaline protease (AKP) as a bioconjugate nano silica-alkaline protease enzyme (BC–SiNP-AKP). This work reports one pot green synthesis of the BC-SiNP-AKP bionanoconjugate complex which included both biotemplating and immobilization of the AKP on to the synthesized silica nanoparticles from cell-free extracts of Bacillus circulans grown in potato peel based medium. Among the cell free crude, acetone concentrated and purified sols of the enzyme AKP, acetone precipitated enzyme sol was found to be best for the biological SiNP synthesis and formation of BC-SiNP-AKP conjugate. The BC-SiNP-AKP had size ranging from 100 to 200 nm with crystalline morphologies varying from spherical, tubular to laminated crystallites. The developed bioconjugate formulation displayed 1.7-fold increase in the enzyme activity post nano-conjugation with superlative dehairing potential on goat skin. The optimized parameters for dehairing were found to be as temperature 37 °C for 24 h of incubation and with enzyme to buffer ratio (2: 50 mL). Thereafter, the dehaired skin was assessed for its histopathological effects, which were found to be safe without any deteriorative changes. The developed formulation is environmentally congenial for its use as depilating agent for animal hides in terms of being green, single pot and cost effective synthesis.