Enzyme immobilization on fibrin

Area of Science:

• Biochemistry
• Biomaterials Science
• Enzyme Engineering

Background:

• Enzyme immobilization is crucial for industrial applications.
• Fibrin offers a versatile matrix for enzyme attachment.
• Understanding immobilization effects on enzyme activity and stability is key.

Purpose of the Study:

• To investigate fibrin as a matrix for immobilizing trypsin.
• To assess the impact of fibrin immobilization on trypsin activity, stability, and kinetics.
• To establish methods for quantifying immobilized enzymes.

Main Methods:

• Covalent immobilization of trypsin onto fibrin powder and membranes.
• Use of Carbon-14 labeled trypsin for quantifying immobilized enzyme.
• Measurement of esterase activity and TAME hydrolysis.
• Assessment of storage stability and substrate inhibition.

Main Results:

• Fibrin successfully immobilized trypsin with retained activity.
• Carbon-14 labeling enabled accurate quantification of immobilized enzyme.
• Enzyme activity was inversely proportional to immobilization levels.
• Enhanced storage stability of immobilized trypsin was observed.
• Optimal activity for TAME hydrolysis was at pH 8-8.4.
• Substrate inhibition occurred above 30mM.

Conclusions:

• Fibrin is a suitable matrix for covalent enzyme immobilization.
• Immobilization affects enzyme kinetics and stability.
• Quantification methods are essential for characterizing immobilized enzyme systems.