IMMOBILIZATION OF CHYMOTRYPSIN ON MAGNETIC Fe3O4 NANOPARTICLES

Lyubov Dishlyuk-,Marina Novoselova-,Tatyana- Rozalyonok-

doi:10.12737/2060

Abstract

Modern methods of chemical modification of enzymes conferring increased catalytic activity and stability to these molecules have been considered. The advantages of using magnetic nanoparticles for the production of stable immobilized enzyme preparations are presented. Chymotrypsin immobilization on Fe3O4 nanoparticles modified with amino groups has been found to result in the incorporation of 88% of the enzyme into the solid phase. The change of the optimal pH and temperature ranges and an increase of stability of the immobilized chymotrypsin relatively to the respective characteristics of the native enzyme have been demonstrated.

Highlights

The possibilities of application of enzymes, especially in medicine and food processing, have expanded significantly due to the recent advances in enzymology
The surface of Fe3O4 nanoparticles is virtually devoid of reactive groups which could be used for covalent binding of the enzyme, and chemical modification prior to enzyme immobilization is necessary
Covalent immobilization of chymotrypsin can be performed using glutaraldehyde which joins the amino groups on the surface of the modified nanoparticles to the amino groups of the enzyme, acting as a spacer

Summary

Introduction

The possibilities of application of enzymes, especially in medicine and food processing, have expanded significantly due to the recent advances in enzymology. The modern methods of enzyme modification confer increased resistance to a variety of chemicals and inhibitors, as well as to pH and temperature effects, to these molecules and allow for alteration of the pH optimum, substrate specificity, and binding properties of the enzymes. The methods of protein engineering based on information on the relationship between amino acid sequence, threedimensional structure, and catalytic activity of enzymes allow for successful modification of enzymes resulting in improvement of their technological characteristics [6, 7].

Results

Conclusion