Lentinula edodes (shiitake mushroom) is the medicinal macro-fungus popular in Japan and China. It is the second most well- known edible fungus in the global market after Agaricus  bisporus Because of presence of dietary fibers, vitamins, proteins, minerals and carbohydrates, shiitake mushroom have significant nutritional worth. It is commonly cultivated in winter season and known as “elixir of life”. Bioactive compounds such as polysaccharides, amino acid derivatives, sterols, glycoproteins, chitin and sulfurous compounds are all extracted from shiitake mushrooms. Some of these are lentinans, eritadenine and KS-2-α- mannan peptide etc. Shiitake mushroom is used therapeutically to cure cancers, cardiovascular disorders, viral and fungal infections, hepatitis, hypertension and hepatic disorders. Shiitake mushroom have also grown in popularity as a result of their unusual and well-liked flavor.

The current study aims to isolate, purify, and characterize the trypsin inhibitor protein from seeds of soya beans, scientifically known as Glycine max. Its seeds were ground, and the powder was soaked several times using n-hexane. It was added to phosphate buffer saline (PBS) followed by filtration and centrifugation of the PBS dissolved extract. The supernatant was subjected to ammonium sulfate precipitation and about six fractions, 30% to 80% were prepared. The centrifuged pellets obtained from each fraction were dialyzed and run on SDS-PAGE. The trypsin inhibitor protein was precipitated and characterized in 30% pellet and molecular weight was 21.5 kDa compared to protein ladder (ThermoFisher 10-170 kDa). GC-MS analysis revealed the steroid derivatives such as stigmasterol, campesterol, beta-sitosterol, and gamma-tocopherol. Glycine max trypsin inhibitor could be used as a plant-derived drug to overcome the over-activation of trypsin without its real substrate (proteins) becoming activated and start auto digestion leading to pancreatitis.