Cellulase from Trichoderma reesei was immobilized by covalent or non-covalent binding onto magnetic hierarchical porous carbon (MHPC) nanomaterials. The immobilization yield and the enzyme activity were higher when covalent immobilization approach was followed. The covalent immobilization approach leads to higher immobilization yield (up to 96%) and enzyme activity (up to 1.35 U mg⁻¹) compared to the non-covalent cellulase binding. The overall results showed that the thermal, storage and operational stability of the immobilized cellulase was considerably improved compared to the free enzyme. The immobilized cellulose catalyzed the hydrolysis of microcrystalline cellulose up to 6 consecutive successive reaction cycles, with a total operation time of 144 h at 50 °C. The half-life time of the immobilized enzyme in deep eutectic solvents-based media was up to threefold higher compared to the soluble enzyme. The increased pH and temperature tolerance of the immobilized cellulase, as well as the increased operational stability in aqueous and deep eutectic solvents-based media indicate that the use of MHPCs as immobilization nanosupport could expand the catalytic performance of cellulolytic enzymes in various reaction conditions.

中文翻译：

---

磁性分层多孔碳纳米粒子上固定的里氏木霉纤维素酶的增强催化性能。

通过共价或非共价结合将里氏木霉的纤维素酶固定在磁性分级多孔碳（MHPC）纳米材料上。当采用共价固定方法时，固定产率和酶活性较高。共价固定化方法导致更高的固定化产率（高达96％）和酶活性（高达1.35 U mg ^-1）与非共价纤维素酶结合。总体结果表明，与游离酶相比，固定化纤维素酶的热稳定性，储存稳定性和操作稳定性得到了显着改善。固定化纤维素催化微晶纤维素的水解，直至连续6个连续反应周期，在50°C下的总操作时间为144 h。与可溶酶相比，在深共熔溶剂基培养基中固定化酶的半衰期最多可延长三倍。固定化纤维素酶的pH值和温度耐受性提高，以及在水性和深共熔溶剂基介质中的操作稳定性提高，表明使用MHPC作为固定化纳米载体可以扩大纤维素酶在各种反应条件下的催化性能。