Rapid screening and isolation of potential xanthine oxidase inhibitors in Ganoderma lucidum

Abstract

Given the abundance and intricate nature of the chemical components found in natural food ingredients, functional food research has long grappled with two significant technical challenges: rapid and precise screening of active substances and targeted preparation. To enhance the efficiency of fungal food preparation, novel methodologies for screening and the production of xanthine oxidase inhibitors from G. lucidum have been developed. The xanthine oxidase inhibitors were rapidly screened using AUF-LC-MS. Molecular docking and molecular dynamics simulation were used to verify the anti-gout activity of active compounds. Subsequently, with activity screening as the guide, HSCCC and SPLC were employed to separate the active ingredients. Simultaneously, the mechanism of action of xanthine oxidase inhibitors was analyzed by enzymatic reaction kinetics. Finally, the anti-gout activity of chemical components in G. lucidum was verified by network pharmacology, and its mechanism of action was discussed. Four potential xanthine oxidase inhibitors, namely ganoderic acid C2, B, A, and D2, were screened from G. lucidum. Their docking binding energies with xanthine oxidase were −6.29, −6.65, −4.29, and −5.96 kcal/mol, respectively. The purity of the four triterpenoid active ingredients isolated from G. lucidum was more than 90 %. These triterpenoid active ingredients showed a reversible inhibitory effect on xanthine oxidase. Network pharmacological studies revealed 71 common targets of active ingredients and gout, obtained by screening and intersecting the two targets, and identified the main metabolic pathways. The results indicate that G. lucidum extract can be employed for the prevention and treatment of gout disease. We enhance the efficiency and purity of active compounds preparation by incorporating enzyme inhibition assays and computerized virtual screening with compound chromatography. With this approach, we seamlessly incorporate fast discovery, screening, and focused preparation of active ingredients, as well as delving into the mechanisms of action of active monomers and proteases, and the molecular-level interactions between active ingredients and proteases. This study providing more opportunities to discover and develop new potential therapeutics from health food resources.

Keywords

• Ganoderma lucidum
• Xanthine oxidase inhibitors
• Chromatography
• Molecular docking
• Network pharmacology