LAUSR

Abstract Agro-food wastes rich in macronutrients are a replacement for expensive carbon and nitrogen sources used for enzyme production. In the present study, we aimed to valorize low-cost agro-wastes for the simultaneous production of protease and lipase haloextremozyme using Haloferax lucentensis GUBF-2 MG076078, characterization of agro-waste residues, and to apply haloextremozyme as Bio-additive in detergent formulation. It was observed that GUBF-2 efficiently hydrolyzed nine different agro-wastes with shrimp waste, and coconut oil cake yielded the highest protease (101.9 U/mL) and lipase (5.83 U/mL), respectively, under optimized conditions of 30% NaCl, pH 6, and 42 °C. Further, SDS-PAGE of cell-free fermented agro-wastes revealed peptides from 212 to 34.6 kDa. Hydrolysates functioned as DPPH radical scavengers with maxima for shrimp waste (45.0 ± 1.15 %). SEM manifested structural alteration of fermented residues. FTIR showed degradation of proteins observed as 1640-1652 and 840-881 cm-1 absorptions with a fading peak at 1742 cm-1 indicating triacylglyceride hydrolysis. Furthermore, haloextremozyme-bio-additive detergent formulation efficiently removed 67.6 ± 3.19% oil and 61.0 ± 3.60% blood stains than 0.5% Triton X-100. Conclusively, Haloferax lucentensis GUBF-2 MG076078 is a prospective strain for simultaneous protease and lipase haloextremozyme production from agro-food wastes, considering both cost efficiency and associated waste disposal problems.