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利用磁性核壳金属有机骨架固定化的猪胰脂肪酶对进行脂肪酶抑制剂的快速筛选。

Rapid Screening of Lipase Inhibitors in by Using Porcine Pancreatic Lipase Immobilized on Magnetic Core-Shell Metal-Organic Frameworks.

机构信息

School of Pharmacy, Shanxi Medical University, 56 Xinjian Road, Taiyuan 030001, China.

出版信息

Molecules. 2022 May 27;27(11):3475. doi: 10.3390/molecules27113475.

DOI:10.3390/molecules27113475
PMID:35684413
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9182405/
Abstract

As for ligand fishing, the current immobilization approaches have some potential drawbacks such as the small protein loading capacity and difficult recycle process. The core-shell metal-organic frameworks composite (FeO-COOH@UiO-66-NH), which exhibited both magnetic characteristics and large specific surface area, was herein fabricated and used as magnetic support for the covalent immobilization of porcine pancreatic lipase (PPL). The resultant composite FeO-COOH@UiO-66-NH@PPL manifested a high loading capacity (247.8 mg/g) and relative activity recovery (101.5%). In addition, PPL exhibited enhanced tolerance to temperature and pH after immobilization. Then, the composite FeO-COOH@UiO-66-NH@PPL was incubated with the extract of to fish out the ligands. Eight lipase inhibitors were obtained and identified by UPLC-Q-TOF-MS/MS. The feasibility of the method was further confirmed through an in vitro inhibitory assay and molecular docking. The proposed ligand fishing technique based on FeO-COOH@UiO-66-NH@PPL provided a feasible, selective, and effective platform for discovering enzyme inhibitors from natural products.

摘要

至于配体钓取，目前的固定化方法存在一些潜在的缺点，如蛋白质负载能力小和回收过程困难。本文制备了具有磁性和大比表面积的核壳型金属有机骨架复合材料（FeO-COOH@UiO-66-NH），并将其用作共价固定猪胰脂肪酶（PPL）的磁性载体。所得的复合 FeO-COOH@UiO-66-NH@PPL 表现出高的负载能力（247.8mg/g）和相对活性回收率（101.5%）。此外，固定化后 PPL 对温度和 pH 值的耐受性增强。然后，将复合 FeO-COOH@UiO-66-NH@PPL 与 的提取物一起孵育以钓取配体。通过 UPLC-Q-TOF-MS/MS 获得并鉴定了 8 种脂肪酶抑制剂。通过体外抑制试验和分子对接进一步证实了该方法的可行性。基于 FeO-COOH@UiO-66-NH@PPL 的配体钓取技术为从天然产物中发现酶抑制剂提供了一种可行、选择性和有效的平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/40504c0880ca/molecules-27-03475-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/2b19f4e0983d/molecules-27-03475-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/f648689db67b/molecules-27-03475-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/653847f9c5b8/molecules-27-03475-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/9793530ea502/molecules-27-03475-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/92468279c65c/molecules-27-03475-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/1439b1acaf92/molecules-27-03475-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/819873c632d8/molecules-27-03475-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/36df7aa2ccca/molecules-27-03475-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/40504c0880ca/molecules-27-03475-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/2b19f4e0983d/molecules-27-03475-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/f648689db67b/molecules-27-03475-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/653847f9c5b8/molecules-27-03475-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/9793530ea502/molecules-27-03475-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/92468279c65c/molecules-27-03475-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/1439b1acaf92/molecules-27-03475-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/819873c632d8/molecules-27-03475-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/36df7aa2ccca/molecules-27-03475-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3085/9182405/40504c0880ca/molecules-27-03475-g009.jpg

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