Rapid Screening and Identification of Antitumor Ingredients from the Mangrove Endophytic Fungus Using an Enzyme-Immobilized Magnetic Nanoparticulate System

Abstract

As a consequence of recent progression in biomedicine and nanotechnology, nanoparticle-based systems have evolved as a new method with extensive applications in responsive therapy, multimodal imaging, drug delivery and natural product separation. Meanwhile, the magnetic nanoparticulate system has aroused great interest for separation and purification because of its excellent magnetic properties. Phospholipase A2 (PLA2) is a highly expressed regulator to promote the growth of various cancers and is an ideal target to treat cancers. In this study, a novel strategy based on ligand–receptor interactions to discover novel PLA2 inhibitors was established, in which PLA2-functionalized Fe3O4@PLGA-PEG-NH2 magnetic nanoparticles were used as a supporting material combined with high-performance liquid chromatography–mass spectrometry, aiming to accelerate the discovery of novel PLA2 inhibitors from natural sources such as mangrove endophytic fungi. Under the optimized ligand fishing conditions, six target compounds were ultimately fished and identified to be cyclic peptides (1–3) and sterols (4–6), which compounds 1, 2 and 4–6 have well-documented cytotoxicities. Compound 3 exerted better inhibitory effect on A549 cells by experiment. In conclusion, PLA2-functionalized Fe3O4@PLGA-PEG-NH2 magnetic nanoparticles-based ligand fishing provided a feasible, selective and effective platform for the efficient screening and identification of antitumor components from natural products.

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

“…The proposed method possesses outstanding advantages over traditional ligand fishing. Apart from the rapidness of microcolumns running, the procedure for MAO‐B immobilization on the microchannel costs only 1.6 hours which is also much faster compared to the 1–2 days routine protein immobilization [43–46]. Secondly, the consumption of protein and herbal samples in the microchannel ligand fishing system is extraordinarily tiny compared to routine ligand fishing (Table S2).…”

On-flow enzymatic inhibitor screening: The emerging success of liquid chromatography-based assays

“…The enzyme immobilization on magnetic particles proved to be a versatile approach in which offline and inline modes could be applied using the same target-support system. Other biomolecules were immobilized by covalent bond through the formation of Schiff bases, like as trypsin (Liu et al, 2019), human Thymidylate Kinase (hTMPK) and the human Nucleoside Diphosphate Kinase (hNDPK) (Ferey et al, 2019), cathepsin D (Cornelio et al, 2018), acetylcholinesterase (AChE) (Vandeput et al, 2015;Seidl et al, 2019), butyrylcholinesterase (BChE) (Seidl et al, 2019), Phospholipase A2 (PLA2) (Wei et al, 2021), and others.…”

Anti-Tumor Secondary Metabolites Originating from Fungi in the South China Sea’s Mangrove Ecosystem

“…A cyclic peptide, fusaristatin C ( 110 ), was rapidly separated from Pseudopithomyces sp. 1512101 and analyzed using a new strategy of ligand fishing based on PLA2-MNPs (Phospholipase A2-functionalized magnetic nanoparticles) with LC–MS (liquid chromatography–mass spectrometry), exerting a potent inhibitory effect on A549 cells, with IC 50 values of 10.10 µM [ 60 ].…”