Isolation of Methyl 5-oxo-DL-prolinate from Agaricus bisporus Mushroom Grown in Natural Conditions and Structural Elucidation Using Spectroscopic Methods

Background: A. bisporus is one of the most important mushrooms in the world for its consumption and valuable chemical composition Aims: This study was conducted to chemically profile secondary metabolites from naturally grown A. bisporus. Study Design: The study adopts a practical experimental design, and the analysis is carried out through structural interpretation of the observed results to derive meaningful conclusions. Place and Duration of Study: The research work was carried out at "Laboratory of Chemistry and Physicochemical Research of Biologically Active Substances" of Karshi State University, March-June 2025. Methodology: In this process, secondary metabolites were isolated from A. bisporus fungus at the chloroform fractionation step by column chromatography, the substances were analyzed by GC-MS and the structure was studied using NMR spectroscopic (H, COSY, ROSY, HBMS, HSQC) methods. Results: At this stage of fractionation, a white amorphous substance was isolated and a mixture of 14 volatile compounds was obtained. They were mainly secondary metabolites belonging to the following classes: complex ester, imide, amine, organic acid, steroids. Between them the amorphous matter was neatly separated. When the chemical structure of the resulting pure substance was determined by physical research methods, it was found that Methyl 5-oxo-DL-prolinate.