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<p>Non-iron magnetic nanoparticles synthesized through a rapid and low-cost process present a promising platform for the oriented immobilization of His-tagged enzymes, enabling efficient magnetic recovery from the reaction medium. In this study, we explored the potential of a novel strategy for developing iron-free magnetic supports aimed at enhancing enzyme reuse in industrial processes. Here, we report the synthesis of cobalt- and nickel-based magnetic nanoparticles via one-pot methods and their evaluation as supports for the targeted immobilization of a recombinant His-tagged β-galactosidase from <i>Kluyveromyces</i> sp. (HisGal). Both supports achieved high immobilization efficiency, with optimal loading at 30 U<sub>enzyme</sub>/g<sub>support</sub>. Structural and compositional analyses, including X-ray diffraction, thermogravimetric analysis, energy-dispersive spectroscopy, and transmission infrared spectroscopy, revealed superior performance of cobalt nanoparticles, enabling more effective oriented immobilization of HisGal. Immobilization on cobalt- or nickel-based nanoparticles shifted the optimal temperature of β-galactosidase to 40&#xa0;°C and 50&#xa0;°C, respectively, and broadened the pH range of enzymatic activity. The immobilized biocatalysts demonstrated enhanced operational stability, maintaining activity over 25 reuse cycles, and improved the resistance of HisGal against thermal degradation and inhibitory effects caused by the presence of sodium, potassium, magnesium, and calcium ions. Moreover, cobalt-based supports significantly reduced galactose inhibition and enhanced lactose affinity in skim milk systems. These findings highlight the versatility and biotechnological potential of iron-free cobalt- and nickel-based magnetic nanoparticles as efficient supports for enzyme immobilization, offering rapid and efficient recovery of immobilized β-galactosidase and contributing to the economic viability of these processes.</p> Graphical Abstract <p></p>

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Iron-free magnetic nanoparticles as a novel support for targeted enzyme immobilization: cobalt- and nickel-based systems

  • 期刊论文

  • 发表日期：

    2025年5月23日

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  • Graziela Paludo,
  • Fernanda Leonhardt,
  • Adriano Gennari,
  • Edilson Valmir Benvenutti,
  • Sabrina Nicolodi,
  • Tânia Maria Haas Costa,
  • Nathalia Denise de Moura Sperotto,
  • Cristiano Valim Bizarro,
  • Luiz Augusto Basso,
  • Pablo Machado,
  • Gaby Renard,
  • Jocelei Maria Chies,
  • Giandra Volpato,
  • Claucia Fernanda Volken de Souza

摘要

Non-iron magnetic nanoparticles synthesized through a rapid and low-cost process present a promising platform for the oriented immobilization of His-tagged enzymes, enabling efficient magnetic recovery from the reaction medium. In this study, we explored the potential of a novel strategy for developing iron-free magnetic supports aimed at enhancing enzyme reuse in industrial processes. Here, we report the synthesis of cobalt- and nickel-based magnetic nanoparticles via one-pot methods and their evaluation as supports for the targeted immobilization of a recombinant His-tagged β-galactosidase from Kluyveromyces sp. (HisGal). Both supports achieved high immobilization efficiency, with optimal loading at 30 Uenzyme/gsupport. Structural and compositional analyses, including X-ray diffraction, thermogravimetric analysis, energy-dispersive spectroscopy, and transmission infrared spectroscopy, revealed superior performance of cobalt nanoparticles, enabling more effective oriented immobilization of HisGal. Immobilization on cobalt- or nickel-based nanoparticles shifted the optimal temperature of β-galactosidase to 40 °C and 50 °C, respectively, and broadened the pH range of enzymatic activity. The immobilized biocatalysts demonstrated enhanced operational stability, maintaining activity over 25 reuse cycles, and improved the resistance of HisGal against thermal degradation and inhibitory effects caused by the presence of sodium, potassium, magnesium, and calcium ions. Moreover, cobalt-based supports significantly reduced galactose inhibition and enhanced lactose affinity in skim milk systems. These findings highlight the versatility and biotechnological potential of iron-free cobalt- and nickel-based magnetic nanoparticles as efficient supports for enzyme immobilization, offering rapid and efficient recovery of immobilized β-galactosidase and contributing to the economic viability of these processes.

Graphical Abstract