Scholarlink

<p>Magnetic nanoparticles functionalized with diethylenetriaminepenta(methylene phosphonic acid) and activated with glutaraldehyde were designed as a heterofunctional support for immobilizing Eversa Transform 2.0 lipase. Structural characterization by X-ray diffraction, transmission and scanning electron microscopies, X-ray fluorescence, Fourier-transform infrared spectroscopy, thermogravimetry, and vibrating-sample magnetometry confirmed preservation of the magnetite spinel phase, spherical primary nanoparticles of approximately 16&#xa0;nm, progressive organic coating, and magnetic responsiveness after enzyme loading. Molecular docking suggested that glutaraldehyde can act as an interfacial bridge between accessible lysine residues of the lipase and the phosphonate-functionalized surface, with calculated interaction energies of −&#xa0;6.0 to −&#xa0;3.6&#xa0;kcal mol<sup>−1</sup> for lysine-glutaraldehyde contacts and −&#xa0;5.4 to −&#xa0;3.2&#xa0;kcal mol<sup>−1</sup> for the phosphonate-glutaraldehyde-lysine complex. A Taguchi L9 design identified protein loading and ionic strength as the most influential immobilization variables, and the optimized condition was 6&#xa0;h, 5 mM buffer, 5&#xa0;mg enzyme per g support, and 40&#xa0;°C.Under these conditions, the immobilized biocatalyst reached 60.5 ± 1.4% immobilization yield and 92.4 ± 2.3% relative activity, corresponding to 94% of the predicted activity. The immobilized enzyme showed no detectable desorption under salt or surfactant challenge, retained more than 50% activity after ten reuse cycles, and preserved approximately 92% of its initial activity after 60 days at 4&#xa0;°C. In solvent-containing assays, immobilization did not universally outperform the soluble enzyme; however, the support improved or maintained tolerance in selected polar organic media, especially acetonitrile and tetrahydrofuran. These results support the material as a magnetically recoverable platform for model ester hydrolysis and future non-aqueous biocatalysis, while direct validation in biodiesel or biolubricant synthesis remains necessary.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Mechanistic and Stability Analysis of Eversa Transform 2.0 Lipase Immobilized on Magnetic Phosphonate-Glutaraldehyde Interfaces

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年6月20日

查看全文
  • Érico Carlos de Castro,
  • Tatiana Sainara Maia Fernandes,
  • Thiago Queiroz da Silva,
  • Paulo Gonçalves de Sousa Junior,
  • Rafael Leandro Fernandes Melo,
  • João Maria Soares,
  • Pierre Basílio Almeida Fechine,
  • Tiago Melo Freire,
  • Antônio Eufrásio Vieira Neto,
  • José Cleiton Sousa dos Santos

摘要

Magnetic nanoparticles functionalized with diethylenetriaminepenta(methylene phosphonic acid) and activated with glutaraldehyde were designed as a heterofunctional support for immobilizing Eversa Transform 2.0 lipase. Structural characterization by X-ray diffraction, transmission and scanning electron microscopies, X-ray fluorescence, Fourier-transform infrared spectroscopy, thermogravimetry, and vibrating-sample magnetometry confirmed preservation of the magnetite spinel phase, spherical primary nanoparticles of approximately 16 nm, progressive organic coating, and magnetic responsiveness after enzyme loading. Molecular docking suggested that glutaraldehyde can act as an interfacial bridge between accessible lysine residues of the lipase and the phosphonate-functionalized surface, with calculated interaction energies of − 6.0 to − 3.6 kcal mol−1 for lysine-glutaraldehyde contacts and − 5.4 to − 3.2 kcal mol−1 for the phosphonate-glutaraldehyde-lysine complex. A Taguchi L9 design identified protein loading and ionic strength as the most influential immobilization variables, and the optimized condition was 6 h, 5 mM buffer, 5 mg enzyme per g support, and 40 °C.Under these conditions, the immobilized biocatalyst reached 60.5 ± 1.4% immobilization yield and 92.4 ± 2.3% relative activity, corresponding to 94% of the predicted activity. The immobilized enzyme showed no detectable desorption under salt or surfactant challenge, retained more than 50% activity after ten reuse cycles, and preserved approximately 92% of its initial activity after 60 days at 4 °C. In solvent-containing assays, immobilization did not universally outperform the soluble enzyme; however, the support improved or maintained tolerance in selected polar organic media, especially acetonitrile and tetrahydrofuran. These results support the material as a magnetically recoverable platform for model ester hydrolysis and future non-aqueous biocatalysis, while direct validation in biodiesel or biolubricant synthesis remains necessary.

Graphical Abstract