Rapid Screening of Lipase Inhibitors in Scutellaria baicalensis by Using Porcine Pancreatic Lipase Immobilized on Magnetic Core–Shell Metal–Organic Frameworks

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 (Fe3O4-COOH@UiO-66-NH2), 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 Fe3O4-COOH@UiO-66-NH2@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 Fe3O4-COOH@UiO-66-NH2@PPL was incubated with the extract of Scutellaria baicalensis 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 Fe3O4-COOH@UiO-66-NH2@PPL provided a feasible, selective, and effective platform for discovering enzyme inhibitors from natural products.

Fast screening of α‐glucosidase inhibitors from Ginkgo biloba leaf by using α‐glucosidase immobilized on magnetic metal‐organic framework

“…Meanwhile, an increasing trend in α‐glucosidase activity was observed within the α‐glucosidase concentration range of 278–1111 µg/mL; however, a decrease in enzyme activity was observed when α‐glucosidase concentration exceeded 1111 µg/mL. This decline may be due to excessive aggregation and agglomeration of enzyme proteins on the carrier surface, resulting in reduced spatial gaps between molecules and hindered interaction between active center structures and substrate molecules [37]. Consequently, 1111 µg/mL was chosen as the optimal immobilization concentration.…”

A rationally engineered specific near-infrared fluorogenic substrate of human pancreatic lipase for functional imaging and inhibitor screening

“…In addition, most of the reported methods involve the use of low-cost porcine pancreatic lipase ( pPL) to replace human pancreatic lipase (hPL) as the enzyme source. 33,34 Although pPL and hPL have high amino acid sequence identity (86%) and a conserved catalytic triad (Asp-His-Ser), species differences in inhibitor responses towards these two lipases have been reported. 35 Consequently, the direct investigation of hPL and high-throughput screening of inhibitors have been realized as practical and reliable methods in complex biological systems.…”

Ultrathin Covalent Organic Framework Nanosheets/Ti3C2Tx-Based Photoelectrochemical Biosensor for Efficient Detection of Prostate-Specific Antigen

“…Therefore, there is an urgent need to develop effective methods to achieve sensitive detection of PSA [ 5 , 6 , 7 , 8 ]. To date, many techniques have been used for the quantitative detection of PSA, including enzyme-linked immunosorbent assay (ELISA), electrochemical technique, colorimetric technique, and fluorescence technique [ 9 , 10 , 11 , 12 ], but the complicated operation and unsatisfactory sensitivity of these methods have greatly hindered their wide application. Therefore, it is essential to explore a reliable, simple, rapid, and sensitive method for prostate-specific antigen detection.…”