Ligand fishing based on tubular microchannel modified with monoamine oxidase B for screening of the enzyme's inhibitors from Crocus sativus and Edgeworthia gardneri

Abstract

A novel strategy of performing ligand fishing with enzyme-modified open tubular microchannel was proposed for screening bioactive components present in medicinal plants. Monoamine oxidase B was immobilized onto the surface of the microchannel for the first time to specifically extract its ligands when the plant's extracts solution flows through the channel. The thermal and the storage stability of immobilized monoamine oxidase B were significantly enhanced after immobilization. Crocin I and II were extracted from Crocus sativus, and tiliroside was extracted from Edgeworthia gardneri. All the three compounds were inhibitors of the enzyme with the half-maximal inhibitory concentration values of 26.70 ± 0.91, 19.88 ± 2.78, and 15.65 ± 0.85 μM, respectively. The enzyme inhibition kinetics and molecular docking were investigated. This is the first report on the inhibitory effects of tiliroside and crocin II. The novel ligand fishing method proposed in this work possesses advantages of rapidness, high efficiency, and tiny sample consumption compared to routine ligand fishing, with promising potential for screening active natural products in complex mixtures.

High-throughput discovery of highly selective reversible hMAO-B inhibitors based on at-line nanofractionation

“…Screening for MAO-B inhibitors from natural products mainly relies on bioassay-guided separation, which often suffers from frequent fractionation and long run times, high costs, and the loss of active substances 10 . To overcome these limitations, magnetic bead ligand fishing 11 and affinity ultrafiltration methods 12 have been proposed, enabling rapid separation of active compound groups and shortening the screening period to several months 13 , 14 , 15 . However, these methods also have limitations, including low throughput and the need for separate analysis after the identification of the active components.…”

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

“…Consequently, ligand-fishing techniques for studying ligand-protein interactions have gained widespread attention [17,18]. This methodology involves the utilization of immobilized target enzymes on carriers to selectively and quickly identify and capture potentially active ligands from complex systems, thereby enhancing the efficiency of screening potentially bioactive natural compounds [19][20][21]. The selection of a carrier material for immobilized enzymes is a critical issue that needs to be addressed.…”

Covalent immobilization of tyrosinase on magnetic multi‐walled carbon nanotubes for inhibitor screening

“…There are some limitations to the use of this method, such as being time-consuming, labor-intensive, and high-cost [11]. At present, ligand fishing technology based on ligandprotein interaction has emerged to improve the efficiency of screening potential active natural compounds [12,13]. In ligand fishing, a target enzyme immobilized on a solid carrier is applied to 'fish' out potential ligands from a complex system.…”