[Native, modified, and immobilized chymotrypsin in chaotropic media. Stabilization limits]

Area of Science:

• Biochemistry
• Protein Stabilization
• Enzyme Engineering

Context:

• Alpha-chymotrypsin is prone to irreversible thermal inactivation at elevated temperatures.
• Developing methods to enhance enzyme stability is crucial for industrial and biotechnological applications.
• Understanding thermal inactivation mechanisms is key to designing effective stabilization strategies.

Purpose:

• To investigate and compare the efficacy of covalent modification, multi-point immobilization, and salting-in compounds in stabilizing alpha-chymotrypsin against thermal inactivation.
• To determine the optimal conditions and combinations of methods for maximizing enzyme stability.
• To elucidate the mechanisms underlying enzyme stabilization.

Summary:

• Various methods, including covalent modification, multi-point immobilization, and salting-in compounds, were employed to stabilize alpha-chymotrypsin at high temperatures.
• Covalent immobilization demonstrated a higher stabilization limit compared to modification or salting-in media alone or in combination.
• Maximum stabilization using salting-in media was achieved with minimal enzyme-support bonds, suggesting suppression of conformational inactivation processes.

Impact:

• Provides insights into effective strategies for enhancing enzyme thermostability.
• Offers potential for improving the performance and longevity of enzymes in industrial processes.
• Contributes to the fundamental understanding of protein inactivation mechanisms and stabilization principles.