Organic-inorganic hybrid nano-materials have been considered to be promising immobilization matrixes for enzymes due to their significantly enhanced reusability and stability of enzymes. Herein, we constructed a novel organic-inorganic hybrid nanoflower via biomacromolecule-regulated biomimetic mineralization to immobilize sucrose phosphorylase (SPase). It was found that chitosan (CS) effectively regulated the biomimetic mineralization of calcium phosphate (CaP), leading to the formation of flower-like hybrid materials for the entrapment of SPase via self-assembly to establish a nano-biocatalyst (CS-CaP@SPase). Upon immobilization, the obtained CS-CaP@SPase exhibited excellent pH, by-product and organic solvents tolerance, and storage stability. Specifically, at acidic condition (pH 4), CS-CaP@SPase performed over 80 % of initial activity, which was 2.42-folds higher than that of free SPase. The catalytic activity of free SPase was severely inhibited about 30 % in the presence of fructose (1.2 M), but CS-CaP@SPase only lost 5 % relative activity. The CS-CaP@SPase retained over 80 % of its relative activity, while the free SPase maintained <20 % of its relative activity in acetonitrile. The relative activity of CS-CaP@SPase was still retained about 80 % after 10 cycles and maintained 75 % after 15 days. Based on Raman spectra analysis, it was also found that the increased β-folding component of SPase in the secondary structure after immobilization was the main factor for its enhanced stability. It is reasonable to believe that biomacromolecule-regulated biomimetic mineralization could be potentially used as a promising method to immobilize enzymes with excellent stability and recyclability, thereby facilitating the preparation of highly efficient catalysts for industrial biocatalysts, biosensing, and biomedicine.

有机-无机杂化纳米材料由于其显着增强的酶的可重复使用性和稳定性而被认为是有前途的酶固定基质。在此，我们通过生物大分子调节的仿生矿化来固定蔗糖磷酸化酶（SPase），构建了一种新型有机-无机杂化纳米花。发现壳聚糖（CS）可有效调节磷酸钙（CaP）的仿生矿化，从而形成花状杂化材料，通过自组装捕获SPase，从而建立纳米生物催化剂（CS-CaP@）斯帕斯）。固定化后，所获得的 CS-CaP@SPase 表现出优异的 pH 值、副产物和有机溶剂耐受性以及储存稳定性。具体来说，在酸性条件（pH 4）下，CS-CaP@SPase 的初始活性超过 80%，比游离 SPase 高 2.42 倍。在果糖（1.2 M）存在下，游离SPase的催化活性被严重抑制约30%，而CS-CaP@SPase的相对活性仅损失5%。CS-CaP@SPase 保留了超过 80% 的相对活性，而游离的 SPase 在乙腈中保持了 <20% 的相对活性。CS-CaP@SPase 的相对活性在 10 个循环后仍保持在 80% 左右，在 15 天后保持在 75%。基于拉曼光谱分析还发现，固定化后二级结构中SPase的β-折叠成分增加是其稳定性增强的主要因素。