Keratin-reinforced encapsulation of whole cells expressing glucose isomerase: Development of robust and reusable biocatalyst microbeads

Abstract

Glucose isomerase (GI) is crucial in high-fructose corn syrup production. This study introduces a novel approach to enhance GI stability and reusability through whole-cell encapsulation of Streptomyces olivochromogenes PTCC 1457 in hybrid microbeads, utilizing keratin as a multifunctional stabilizer and cross-linker. Optimal bead formation was achieved using 2 % alginate, 2-3 % CaCl₂, and 2.5 % keratin at pH 7.0 and 37-40 °C. Keratin played a vital role in forming a robust and flexible matrix. Immobilization in keratin-alginate-biomass beads maintained GI activity (655 GIU·g^-1) comparable to free enzyme (650 GIU·g^-1), while silicate incorporation reduced activity to 234 GIU·g^-1. The immobilized enzyme exhibited enhanced stability over a wider pH (6-9) and temperature (4-60 °C) range compared to the free enzyme. Importantly, the immobilized GI maintained 80 % of its initial activity after 20 reaction cycles. Thermogravimetric analysis, scanning electron microscopy, energy dispersive X-ray spectroscopy, and tensile testing confirmed the formation of hybrid beads with improved thermal and mechanical stability. This novel immobilization strategy, leveraging keratin's unique properties, offers a promising approach for enhancing GI stability, reusability, and storage longevity, potentially improving its industrial applicability in high-fructose corn syrup production.

Keywords: Glucose isomerase; Keratin; Whole-cell encapsulation.