Immobilized chymotrypsin on reversibly precipitable polymerized liposome

Area of Science:

• Biotechnology
• Materials Science
• Enzyme Engineering

Background:

• Polymerized liposomes (PLS) offer a unique platform for biomolecule immobilization due to their tunable properties.
• Enzyme immobilization is crucial for enhancing enzyme stability, reusability, and process efficiency in biocatalysis.

Purpose of the Study:

• To develop a novel method for covalently immobilizing chymotrypsin onto the surface of synthesized polymerized liposomes (PLS).
• To evaluate the activity, stability, and reusability of the chymotrypsin-immobilized PLS conjugate.

Main Methods:

• Synthesis of a phosphatidylethanolamine with a diacetylene moiety for PLS formation.
• Covalent immobilization of chymotrypsin onto PLS using the carbodiimide method.
• Activity assays using low- and high-molecular-weight substrates (N-benzoyl-L-tyrosine ethyl ester and casein).

Main Results:

• Efficient and rapid enzyme coupling achieved at a low enzyme-to-PLS weight ratio (< 0.12).
• High activity yields observed for both small (90 +/- 9%) and large (59 +/- 5%) substrates.
• Immobilized enzyme demonstrated superior stability at high temperatures and during prolonged incubation compared to native chymotrypsin, along with excellent reusability.

Conclusions:

• Covalent immobilization of chymotrypsin onto PLS via the carbodiimide method is a highly effective strategy.
• The resulting immobilized enzyme exhibits enhanced operational stability, reusability, and broad substrate specificity.
• This approach holds significant potential for applications in biocatalysis and enzyme-based technologies.