School of Pharmacy, Xi'an Jiaotong University, Xi'an 710061, China.
Analyst. 2019 Dec 16;145(1):268-276. doi: 10.1039/c9an01777a.
Immobilized enzymes play significant roles in many practical applications. However, the enzymes need to be purified before immobilization by conventional immobilizing methods, and the purification process is expensive, laborious, complicated and results in a decrease of the enzymatic activity. So, we present a novel method by a facile one-step targeted immobilization of an enzyme without a purification process from complex samples. For this purpose, a novel molecularly imprinted polymer was prepared via a silane emulsion self-assembly method using boric acid-modified Fe3O4 nanoparticles as magnetic nuclei, horseradish peroxidase as a template, 3-aminopropyltriethoxysilane as a functional monomer and tetraethyl orthosilicate as a crosslinking agent. The molecularly imprinted polymers were characterized using a scanning electron microscope, X-ray photoelectron spectroscope, vibrating sample magnetometer and X-ray diffractometer. The as-prepared and characterized materials were employed to immobilize horseradish peroxidase from a crude extract of horseradish. Moreover, the immobilized horseradish peroxidase was employed to develop visual sensors for the detection of glucose and sarcosine. This study demonstrated that the molecularly imprinted polymers prepared via the silane emulsion self-assembly method can facilely immobilize horseradish peroxidase from a crude extract of horseradish without any purification process. The developed visual method based on the immobilized horseradish peroxidase shows great potential applications for the visual detection of glucose and sarcosine.
固定化酶在许多实际应用中起着重要作用。然而，传统的固定化方法需要在固定化之前对酶进行纯化，而纯化过程昂贵、繁琐、复杂，并且会导致酶活性降低。因此，我们提出了一种新的方法，通过简便的一步靶向固定化，从复杂的样品中无需纯化过程即可获得酶。为此，我们通过使用硼酸改性的 Fe3O4 纳米粒子作为磁性核、辣根过氧化物酶作为模板、3-氨丙基三乙氧基硅烷作为功能单体和正硅酸乙酯作为交联剂的硅烷乳液自组装方法制备了一种新型的分子印迹聚合物。使用扫描电子显微镜、X 射线光电子能谱仪、振动样品磁强计和 X 射线衍射仪对分子印迹聚合物进行了表征。所制备的材料经过表征后，用于从辣根粗提物中固定辣根过氧化物酶。此外，固定化辣根过氧化物酶用于开发用于检测葡萄糖和肌氨酸的可视化传感器。这项研究表明，通过硅烷乳液自组装方法制备的分子印迹聚合物可以简便地从辣根粗提物中固定辣根过氧化物酶，而无需任何纯化过程。基于固定化辣根过氧化物酶的开发的可视化方法在可视化检测葡萄糖和肌氨酸方面具有很大的应用潜力。
