Covalent Lysozyme Immobilization on Enzymatic Cellulose Nanocrystals

Chemistry. 2024 Oct 28;30(60):e202402171. doi: 10.1002/chem.202402171. Epub 2024 Oct 16.

Abstract

Nanostructured materials represent promising substrates for biocatalyst immobilization and activation. Cellulose nanocrystals (CNCs), accessible from waste and/or renewable sources, are sustainable and biodegradable, show high specific surface area for anchoring a high number of enzymatic units, and high thermal and mechanical stability. In this work, we present a holistic enzyme-based approach to functional antibacterial materials by bioconjugation between the lysozyme from chicken egg white and enzymatic cellulose nanocrystals. The neutral CNCs were prepared by endoglucanase hydrolysis from Avicel. We explore the covalent immobilization of lysozyme on enzymatic CNCs and on their TEMPO oxidized derivatives (TO-CNCs), comparing immobilization yields, material properties, and enzymatic activities. The materials were characterized by X-ray diffractometry (XRD), attenuated total reflectance Fourier Transform infrared spectroscopy (ATR-FTIR), bicinchoninic acid (BCA) assay, field-emission scanning electron microscopy (FE-SEM) and dynamic light scattering (DLS). We demonstrate the higher overall efficiency of the immobilization process carried out on TO-CNCs, based on the success of covalent bonding and on the stability of the isolated bioconjugates.

Keywords: Biocatalysis; Bioconjugation; Cellulose nanocrystals; Enzymes immobilization; Lysozyme.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / chemistry
  • Cellulose* / chemistry
  • Chickens
  • Cyclic N-Oxides / chemistry
  • Enzymes, Immobilized* / chemistry
  • Enzymes, Immobilized* / metabolism
  • Hydrolysis
  • Muramidase* / chemistry
  • Muramidase* / metabolism
  • Nanoparticles* / chemistry
  • Spectroscopy, Fourier Transform Infrared
  • X-Ray Diffraction

Substances

  • Cellulose
  • Muramidase
  • Enzymes, Immobilized
  • TEMPO
  • Cyclic N-Oxides
  • Anti-Bacterial Agents