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Carboxy-Functionalized Covalent Organic Framework as a Carrier for Lipase Immobilization and Its Application in Inhibitors Screening

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Abstract

Covalent organic frameworks (COFs) with large specific surface areas, high porosity, good stability, and designable structure are promising carriers for immobilized enzymes. It is important to explore lipase inhibitors from natural foods as lipase inhibitors are closely related to the treatment of obesity. In this work, a carboxyl functionalized covalent organic framework (TpBD-3COOH) was prepared by solvothermal method for covalent immobilization of porcine pancreatic lipase (PPL) and obtained the enzyme-decorated COF (PPL@COF). The immobilized lipase showed wider pH and temperature tolerance with the same optimal pH and temperature of 7.5 and 50 ℃ compared to free lipase. After 6 successive reuses, the PPL@COF maintained 53.0% of its original activity. Immobilized lipase also displayed enhanced storage stability (55.4% after 14 days at 4 ℃). When p-nitrophenyl acetate was applied as the substrate, the calculated Michaelis constant was 3.57 mM and the half maximal inhibitory concentration of orlistat was 3.20 μM. Finally, the PPL@COF was used for enzyme inhibitors screening from natural foods combined with UV spectrophotometry, and Hawthorn was screened for excellent lipase inhibitory activity.

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Funding

This work was supported by the Natural Science Foundation of Jiangsu Province (BE2019717).

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Authors and Affiliations

  1. Department of Analytical Chemistry, China Pharmaceutical University, 24 TongJiaXiang, Nanjing, 210009, Jiangsu, China

    Xue Liu, Jiaqi Wu, Shan Yang, Lingyu Li & Yibing Ji

  2. Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, Nanjing, 210009, China

    Xue Liu, Jiaqi Wu, Shan Yang, Lingyu Li & Yibing Ji

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  1. Xue Liu
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  3. Shan Yang
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Contributions

Yibing Ji contributed to the conception of the study; Xue Liu completed the experiments, collected the data and wrote the manuscript; Jiaqi Wu helped provide the analysis with constructive discussions and check the paper; Shan Yang and Lingyu Li helped charting. All authors read and approved the final manuscript.

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Liu, X., Wu, J., Yang, S. et al. Carboxy-Functionalized Covalent Organic Framework as a Carrier for Lipase Immobilization and Its Application in Inhibitors Screening. Appl Biochem Biotechnol (2023). https://doi.org/10.1007/s12010-023-04725-1

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