[Some physicochemical properties of modified trypsin].

Area of Science:

• Biochemistry
• Enzymology
• Protein Chemistry

Context:

• Investigating enzyme modification and immobilization techniques.
• Understanding the stability and activity of enzymes in complex biological matrices.
• Exploring the use of glutaraldehyde as a cross-linking agent for protein conjugation.

Purpose:

• To study the physico-chemical properties of trypsin covalently bound to human serum albumin (HSA) using glutaraldehyde.
• To assess the impact of this modification on trypsin's pH optimum and susceptibility to inhibitors.
• To evaluate the effect of the modification on the rate of autolytic degradation.

Summary:

• Physico-chemical properties of glutaraldehyde-crosslinked trypsin-HSA conjugate were analyzed.
• The modification minimally affected the pH optimum of trypsin activity.
• Inhibition studies with soybean and human serum inhibitors were performed, yielding apparent inhibition constants.
• Crucially, the modification significantly decelerated the autolytic degradation of trypsin, with autolysis rate constants determined at 50°C.

Impact:

• Provides insights into stabilizing enzymes through conjugation with serum albumin.
• Demonstrates a method to enhance enzyme resistance to autolysis, potentially increasing enzyme shelf-life and utility.
• Offers valuable data for applications requiring robust and stable trypsin preparations.