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
{"title":"Iron-free magnetic nanoparticles as a novel support for targeted enzyme immobilization: cobalt- and nickel-based systems","authors":"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","doi":"10.1007/s10562-025-05057-x","DOIUrl":null,"url":null,"abstract":"<div><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 °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.</p><h3>Graphical Abstract</h3><div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":508,"journal":{"name":"Catalysis Letters","volume":"155 6","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Catalysis Letters","FirstCategoryId":"92","ListUrlMain":"https://link.springer.com/article/10.1007/s10562-025-05057-x","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, PHYSICAL","Score":null,"Total":0}
引用次数: 0
Abstract
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.
期刊介绍：
Catalysis Letters aim is the rapid publication of outstanding and high-impact original research articles in catalysis. The scope of the journal covers a broad range of topics in all fields of both applied and theoretical catalysis, including heterogeneous, homogeneous and biocatalysis.
The high-quality original research articles published in Catalysis Letters are subject to rigorous peer review. Accepted papers are published online first and subsequently in print issues. All contributions must include a graphical abstract. Manuscripts should be written in English and the responsibility lies with the authors to ensure that they are grammatically and linguistically correct. Authors for whom English is not the working language are encouraged to consider using a professional language-editing service before submitting their manuscripts.
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