Functional stabilization of trypsin by conjugation with beta-cyclodextrin-modified carboxymethylcellulose

Area of Science:

• Biochemistry
• Enzyme Engineering
• Polymer Chemistry

Background:

• Bovine trypsin is a widely used protease with applications in various industries.
• Native trypsin can be susceptible to denaturation and autolysis, limiting its operational stability.
• Chemical modification offers a route to improve enzyme properties for industrial applications.

Purpose of the Study:

• To chemically conjugate bovine pancreatic trypsin with a beta-cyclodextrin-carboxymethylcellulose polymer.
• To evaluate the impact of this polymer conjugation on trypsin's enzymatic activity, thermal stability, and resistance to degradation.

Main Methods:

• Bovine trypsin was modified using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as a coupling agent.
• Enzymatic activity (esterolytic and proteolytic) was measured before and after conjugation.
• Thermal stability, thermostability, and resistance to autolysis and sodium dodecyl sulfate denaturation were assessed.

Main Results:

• The trypsin-polymer conjugate retained high levels of esterolytic (110%) and proteolytic (95%) activity.
• Conjugation increased the optimal temperature by 8°C and thermostability by 16°C.
• The modified trypsin exhibited enhanced stability against thermal incubation, autolysis at pH 9.0, and sodium dodecyl sulfate denaturation.

Conclusions:

• Chemical modification of bovine trypsin with a beta-cyclodextrin-carboxymethylcellulose polymer significantly enhances its functional properties.
• The resulting polymer-enzyme complex demonstrates improved stability and activity, making it suitable for broader applications.
• This approach offers a viable strategy for developing more robust and stable enzyme preparations.