Cloning and expression of a chimeric protease and chitinase fusion protein from Metarhizium anisopliae into Escherichia coli for enhanced enzyme activity

Abstract

This thesis focuses on developing chimeric protease, chitinase, and protease-chitinase fusion proteins with increased binding affinity to insect cuticles and improved catalytic activity to enhance cuticle degradation. The research explores the insecticidal capabilities of these engineered enzymes, aiming to combat insect pests more effectively. By combining the wild-type Pr1a protease and Chi2 chitinase from Metarhizium anisopliae with a chitin-binding domain from Bombyx mori, the engineered proteins are designed to breach the insect's primary defense mechanism—the cuticle—more rapidly. Enhancing these enzymes' efficiency directly increases fungal virulence, making it a more potent pathogen.