Integrated In Silico-In Vitro Analysis of Amylase Production from Agro-Waste by Bacillus subtilis with Molecular Docking Studies of Enzyme-Substrate Interactions

Abstract

Sustainable enzyme production from agro-industrial residues presents significant opportunities for cost-effective bioprocess development. This study integrates experimental fermentation and computational modeling to optimize amylase production by Bacillus subtilis using agro-waste substrates. Submerged fermentation parameters, including substrate concentration, pH, temperature, and incubation time, were structured for optimization to enhance enzyme yield. Enzyme activity quantification and partial purification protocols were incorporated to evaluate catalytic performance. Concurrently, in silico structural modeling and docking analyses were performed to investigate enzyme-substrate binding interactions and active-site architecture. Interaction mapping aimed to correlate structural insights with catalytic efficiency and substrate specificity. The integrated framework seeks to combine empirical fermentation data with computational mechanistic understanding to guide process optimization. This approach supports sustainable enzyme production strategies while advancing rational bioprocess engineering methodologies.