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<p>Sucrose isomerase (SIM) faces challenges in industrial applications due to its limited stability and reusability. To address these issues, a copper ion-mediated hybrid nanoflower immobilization system was developed for a rationally engineered SIM mutant (Q310E). Under optimized synthesis conditions (50&#xa0;h, 120 mM Cu<sup>2+</sup>, SIM-to-Cu<sup>2+</sup> ratio of 0.012&#xa0;mg/mM, 4℃), the resulting SIM@Cu-NFs exhibited a well-defined hierarchical structure and high batch-to-batch reproducibility. The immobilized enzyme showed significantly enhanced stability, retaining over 80% activity after 1&#xa0;h at pH 4.0–8.0, over 70% activity after 3&#xa0;h at 50 ℃, and over 70% activity after 45 days of storage at 4℃. Moreover, it maintained 55.9% activity after 6 reuse cycles and approximately 45% after 12 cycles. Kinetic analysis revealed a 38.5% increase in catalytic efficiency (<i>K</i><sub>cat</sub>/<i>K</i><sub>m</sub>), along with improved substrate affinity (<i>K</i><sub>m</sub> decreased from 53.5 to 45.0 ± 0.7 mM) and a higher reaction rate (<i>V</i><sub>max</sub> increased from 1609 to 1950 ± 28 µM·min<sup>− 1</sup>). These enhancements are attributed to the stabilizing effect of Cu<sup>2+</sup> coordination bonds and the favorable microenvironments within the nanoflower architecture. The SIM@Cu-NF system demonstrates high operational stability, reusability, and catalytic efficiency, showing promising potential for industrial applications in enzyme-based biocatalysis.</p>

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Enhanced Catalytic Performance and Stability of Sucrose Isomerase Via Cu2+-Mediated Nanoflower Immobilization

  • 期刊论文

  • 发表日期：

    2026年1月21日

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  • Tiantian Gong,
  • Jianing Zhang,
  • Caifeng Li,
  • Huilian Xu,
  • Jinsong Gu

摘要

Sucrose isomerase (SIM) faces challenges in industrial applications due to its limited stability and reusability. To address these issues, a copper ion-mediated hybrid nanoflower immobilization system was developed for a rationally engineered SIM mutant (Q310E). Under optimized synthesis conditions (50 h, 120 mM Cu2+, SIM-to-Cu2+ ratio of 0.012 mg/mM, 4℃), the resulting SIM@Cu-NFs exhibited a well-defined hierarchical structure and high batch-to-batch reproducibility. The immobilized enzyme showed significantly enhanced stability, retaining over 80% activity after 1 h at pH 4.0–8.0, over 70% activity after 3 h at 50 ℃, and over 70% activity after 45 days of storage at 4℃. Moreover, it maintained 55.9% activity after 6 reuse cycles and approximately 45% after 12 cycles. Kinetic analysis revealed a 38.5% increase in catalytic efficiency (Kcat/Km), along with improved substrate affinity (Km decreased from 53.5 to 45.0 ± 0.7 mM) and a higher reaction rate (Vmax increased from 1609 to 1950 ± 28 µM·min− 1). These enhancements are attributed to the stabilizing effect of Cu2+ coordination bonds and the favorable microenvironments within the nanoflower architecture. The SIM@Cu-NF system demonstrates high operational stability, reusability, and catalytic efficiency, showing promising potential for industrial applications in enzyme-based biocatalysis.