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脯氨酸修饰的 UIO-66 作为纳米载体,提高脂肪酶的催化活性和稳定性,用于电化学检测硝基酚。

Proline-Modified UIO-66 as Nanocarriers to Enhance Lipase Catalytic Activity and Stability for Electrochemical Detection of Nitrofen.

机构信息

Department of Food Science and Engineering, College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315211, China.

Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition, Ningbo 315211, China.

出版信息

ACS Appl Mater Interfaces. 2021 Jan 27;13(3):4146-4155. doi: 10.1021/acsami.0c17134. Epub 2021 Jan 13.

Abstract

Immobilization can be used to improve the stability of lipases and enhances lipase recovery and reusability, which increases its commercial value and industrial applications. Nevertheless, immobilization frequently causes conformational changes of the lipases, which decrease lipase catalytic activity. in the present work, we synthesized UIO-66 and grafted UIO-66 crystals with proline for immobilization of lipase (CRL). As indicated by steady-state fluorescence microscopy, grafting of proline onto UIO-66 crystals induced beneficial conformational change in CRL. CRL immobilized on UIO-66/Pro (CRL@UIO-66/Pro) demonstrated higher enzyme activity and better recyclability than that immobilized on UIO-66 (CRL@UIO-66) in both hydrolysis (CRL@UIO-66/Pro: 0.34 U; CRL@UIO-66: 0.15 U) and transesterification (CRL@UIO-66/Pro: 0.93 U; CRL@UIO-66: 0.25 U) reactions. The higher values of and / of CRL@UIO-66/Pro also showed that it had better catalytic efficiency as compared to CRL@UIO-66. It is also worth noting that CRL@UIO-66/Pro (0.93 U) demonstrated a much higher transesterification activity as compared to free CRL (0.11 U), indicating that UIO-66/Pro has increased the solvent stability of CRL. Both CRL@UIO-66 and CRL@UIO-66/Pro were also used for the fabrication of biosensors for nitrofen with a wide linear range (0-100 μM), lower limit of detection, and good recovery rate.

摘要

固定化可以提高脂肪酶的稳定性,并增强脂肪酶的回收和重复使用性,从而提高其商业价值和工业应用。然而,固定化经常导致脂肪酶构象发生变化,从而降低脂肪酶的催化活性。在本工作中,我们合成了 UIO-66,并将其接枝到脯氨酸上,用于固定化脂肪酶(CRL)。稳态荧光显微镜表明,脯氨酸接枝到 UIO-66 晶体上诱导了 CRL 的有利构象变化。与固定在 UIO-66 上的 CRL(CRL@UIO-66)相比,固定在 UIO-66/Pro(CRL@UIO-66/Pro)上的 CRL 在水解(CRL@UIO-66/Pro:0.34 U;CRL@UIO-66:0.15 U)和转酯(CRL@UIO-66/Pro:0.93 U;CRL@UIO-66:0.25 U)反应中表现出更高的酶活性和更好的可重复使用性。CRL@UIO-66/Pro 的 值和 / 值也更高,表明其催化效率优于 CRL@UIO-66。值得注意的是,与游离 CRL(0.11 U)相比,CRL@UIO-66/Pro 具有更高的转酯活性(0.93 U),表明 UIO-66/Pro 提高了 CRL 的溶剂稳定性。CRL@UIO-66 和 CRL@UIO-66/Pro 都被用于制备对硝基酚的生物传感器,具有较宽的线性范围(0-100 μM)、更低的检测限和良好的回收率。

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