Chitosan–Collagen Coated Magnetic Nanoparticles for Lipase Immobilization—New Type of “Enzyme Friendly” Polymer Shell Crosslinking with Squaric Acid

Marta Ziegler-Borowska,Katarzyna Wegrzynowska-Drzymalska,Adam Sikora,Dorota Chelminiak-Dudkiewicz,Tomasz Siódmiak,Joanna Skopinska-Wisniewska,Halina Kaczmarek,Michał Marszałł

doi:10.3390/catal7010026

Marta Ziegler-Borowska, Katarzyna Wegrzynowska-Drzymalska + Show 6 more

Open Access

https://doi.org/10.3390/catal7010026

Copy DOI

Abstract

This article presents a novel route for crosslinking a polysaccharide and polysaccharide/protein shell coated on magnetic nanoparticles (MNPs) surface via condensation reaction with squaric acid (SqA). The syntheses of four new types of collagen-, chitosan-, and chitosan–collagen coated magnetic nanoparticles as supports for enzyme immobilization have been done. Structure and morphology of prepared new materials were characterized by attenuated total reflectance Fourier-transform infrared (ATR-FTIR), XRD, and TEM analysis. Next, the immobilization of lipase from Candida rugosa was performed on the nanoparticles surface via N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC)/N-hydroxy-succinimide (NHS) mechanism. The best results of lipase activity recovery and specific activities were observed for nanoparticles with polymer shell crosslinked via a novel procedure with squaric acid. The specific activity for lipase immobilized on materials crosslinked with SqA (52 U/mg lipase) was about 2-fold higher than for enzyme immobilized on MNPs with glutaraldehyde addition (26 U/mg lipase). Moreover, a little hyperactivation of lipase immobilized on nanoparticles with SqA was observed (104% and 112%).

Highlights

Lipases are one of the most used hydrolases, and are applied in wide range of organic reactions as well as in analytical procedures [1,2]
Six types of magnetic nanoparticles (MNPs) coated with chitosan, collagen, and the blend of these two biopolymers were prepared by standard coprecipitation reaction
Two types of crosslinkers were used for polymer layer stabilization: glutaraldehyde (Glu) and squaric acid (SqA) (Figure 1)

Summary

Introduction

Lipases are one of the most used hydrolases, and are applied in wide range of organic reactions as well as in analytical procedures [1,2]. The active center of the lipases from Candida rugosa is covered by a polypeptide chain called lid. This polypeptide can rearrange and leads to conformational changes of lipases at the interface of water and organic phases. One of the most promising methods to improve the enzyme’s disadvantages is immobilization [7,8]. The strategy of immobilization has a great impact on the attached enzyme properties—it may improve enzyme activity, stability, selectivity, and specificity [9,10]

Methods

Results

Conclusion