Harnessing the Proteolytic Power of Aspergillus Terreus Sgtm2

Objectives: The research is focused on isolating and characterizing proteolytic microorganisms from soil samples collected from a cowshed. Additionally, it aimed to optimize the conditions for protease production by utilizing a selected microbial strain. Methods: Cowshed soil were used to isolate protease-producing fungi; they were cultured on PDA (Potato Dextrose Agar) plates and tested for their ability to produce proteolytic enzymes using milk agar plates. Morphological analysis and 18S rRNA sequencing identified the strains. Submerged fermentation was employed for protease production, with quantification conducted using the McDonald and Chen assay method. Process optimization was performed to maximize enzyme yield under varying conditions of inoculum size, temperature, pH, carbon, and nitrogen sources. Findings: Among five isolates (C1, C2, C3, C4, and C5), four exhibited proteolytic activity, with isolate C3 showing the highest protein hydrolysis. Morphological and molecular analysis identified the most efficient strain, SGTM2, as Aspergillus terreus with 99.10% similarity to Aspergillus terreus ATE1. Optimized production conditions (2% inoculum size, 45°C, pH 8.0, 1% maltose as carbon source, and 1.5% peptone as nitrogen source) resulted in a yield of 5.03 kg/m³ protease, with peak production achieved on the 4th day of incubation. Novelty: This study demonstrates the industrial potential of Aspergillus terreus in producing high yields of protease under optimized conditions. The study shows that the species differ 0.9% from the existing species which leads to the better and high yield of protease (accession number PP474258). The study contributes to the development of cost-effective enzyme production processes, particularly for applications in detergents, food processing, pharmaceuticals, and waste management industries. Keywords: Protease synthesis, Aspergillus terreus, Bio waste management enzymes, Protein hydrolysis, Cowshed soil fungi, Soil-derived fungi