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

IF 3.1 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Applied Biochemistry and Biotechnology Pub Date : 2023-10-11 DOI:10.1007/s12010-023-04725-1

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

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引用次数: 0

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.

Graphical Abstract

Abstract Image

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羧官能化共价有机骨架作为脂肪酶固定化载体及其在抑制剂筛选中的应用。

共价有机框架（COFs）具有高比表面积、高孔隙率、良好的稳定性和可设计的结构，是固定化酶的理想载体。从天然食品中发掘脂肪酶抑制剂具有重要意义，因为脂肪酶抑制剂与肥胖的治疗密切相关。本工作采用溶剂热法制备了羧基功能化共价有机骨架（TpBD-3COOH），用于共价固定化猪胰脂肪酶（PPL），并获得了酶修饰的COF(PPL@COF)。与游离脂肪酶相比，固定化脂肪酶在相同的最适pH和温度7.5和50℃下表现出更宽的pH和温度耐受性。在连续6次重复使用后PPL@COF维持了原来活性的53.0%。固定化脂肪酶的贮藏稳定性也有所提高（在4℃条件下贮藏14天后提高55.4%）。当使用对硝基苯基乙酸酯作为底物时，计算的米氏常数为3.57mM，奥利司他的半最大抑制浓度为3.20μM。最后PPL@COF采用紫外分光光度法从天然食品中筛选酶抑制剂，筛选出山楂具有良好的脂肪酶抑制活性。

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来源期刊

Applied Biochemistry and Biotechnology 工程技术-生化与分子生物学

CiteScore

5.70

自引率

6.70%

发文量

460

审稿时长

5.3 months

期刊介绍： This journal is devoted to publishing the highest quality innovative papers in the fields of biochemistry and biotechnology. The typical focus of the journal is to report applications of novel scientific and technological breakthroughs, as well as technological subjects that are still in the proof-of-concept stage. Applied Biochemistry and Biotechnology provides a forum for case studies and practical concepts of biotechnology, utilization, including controls, statistical data analysis, problem descriptions unique to a particular application, and bioprocess economic analyses. The journal publishes reviews deemed of interest to readers, as well as book reviews, meeting and symposia notices, and news items relating to biotechnology in both the industrial and academic communities. In addition, Applied Biochemistry and Biotechnology often publishes lists of patents and publications of special interest to readers.