A Novel Approach to Enhancing Protein Stability through Site-Directed Mutagenesis and Computational Design

• Integration of molecular dynamics and machine learning enables accurate prediction of stabilizing mutations. • Engineered variants showed up to 15°C increase in melting temperature (Tm) and enhanced resistance to chemical denaturation. • The approach is generalizable and can be applied to various enzymes and therapeutic proteins. • The engineered enzyme retained catalytic activity under harsh industrial conditions, demonstrating practical utility.