Characterization of Secondary Metabolites Identified in Lentinula edodes and Determination of Their Biological Activities by in‐silico Antioxidant Screening

Abstract Mushrooms, like other fungi, are unique in the living world They have been assigned to the kingdom of Myceteae. Lentinula edodes (shiitake), is second‐largest and most often consumed edible mushroom in the world. Shiitake mushroom possess large capability to be used as food as it contains carbohydrates, proteins, vitamins, minerals, fibers, fatty acids, amino acids, etc. In current study powdered form mushroom used to form semisolid extract in selected fractions (Butanol, ethyl acetate, hexane, chloroform and aqueous). The aim of the current study was to explore phytoconstituents which have biological activities in selected fractions (n. hexane chloroform, and ethyl acetate) of methanolic extract of Lentinula edodes .GC‐MS, and HPLC analysis presented the secondary metabolites in all the selected fractions under taken. Chloroform fraction had the highest total phenolic, and total flavonoid which exhibited the maximum in vitro antioxidant ABTS potential (74.78±0.83) potential at 350 μg/mL while ethyl acetate has moderate and n‐hexane shows least concentration. Hemolysis essay showed less than 10 % lysis of RBC, s which indicates that fractions under study are active, and safe to use as herbal medicine. Biomolecules detected through HPLC and GC‐MS in fractions under study showed significant inhibition in paw inflammation (1.18±0.06) and yeast induced pyrexia (36.8±0.68) in dose dependent manner. Furthermore, In‐silico study showed that 4H‐Furo[3,2‐b] pyrrole‐5‐carboxylic acid have a strong interaction to NADPH oxidase, Myeloperoxidase as compared to standard Melatonin which shows best antioxidant compound which was confirmed in chloroform fraction.The outcomes of this research indicated that L. edodes significantly reduced inflammation and have ability to scavenge free radicals, prevent protein denaturation, also indicate the possible use in food and nutraceutical applications as an antioxidant, antipyretic, anti‐inflammatory, and antidiabetic agent.