Pectin nanoparticles as a biodegradable support for d-tagatose 3-epimerase immobilization.

Abstract

Rare sugars have attracted increasing interest because of their potential health benefits, although industrial production is often limited by the poor stability and reusability of free enzymes. In this study, d-tagatose 3-epimerase (DTE), a key enzyme for the production of rare sugar d-sorbose, was immobilized on synthesized pectin nanoparticles (PNPs) through covalent bonding using carbodiimide-mediated coupling chemistry. Successful immobilization was confirmed by physicochemical characterization using techniques, namely Dynamic Light Scattering, Field Emission Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy, and Fluorescence Microscopy. The resulting immobilized DTE (PNP@DTE) exhibited an average particle size of 585.7 ± 15 nm with a Polydispersity Index (PDI) of 0.252 ± 0.018. The developed system showed an enzyme loading of about 75 ± 3% and an activity recovery of 79 ± 2%. PNP@DTE retained 90% of its initial activity after 30 days of storage at 4 °C. It also demonstrated good pH and thermal stability, retaining approximately 90% residual activity after 60 min at 50 °C and over 85% activity after 12 h at pH 8-9. Furthermore, the PNP@DTE maintained nearly 80% of its initial activity after six consecutive reuse cycles. These findings demonstrate the feasibility of using pectin nanoparticles as an alternative biopolymer-based support for DTE immobilization for the sustainable production of d-sorbose.

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