Presence of Protease Inhibitors in Agaricus blazei Murril Mushrooms

Brazil stands as the largest producer of Agaricus blazei, being a native species, with favorable climatic conditions for its cultivation. Thus, it stands out as an object of study on the nutritional properties and pharmacological features. Allegedly, Agaricus blazei is responsible for some beneficial properties to human health, such as immunomodulatory activity, antioxidant, anti-inflammatory and anti-cancer, and has been used to treat certain diseases. The aim was to evaluate the presence of protease inhibitors in five Agaricus extracts. The extracts of Agaricus blazei using methanol, ethanol, butanol, propanol and water (after lyophilization) were solubilized in Hepes buffer to a final concentration of 5mg/ml. The enzymatic method was used for analysis of samples. Benzoyl-DL-arginine-p-nitroanilide (BApNA) was used as substrate in a solution using 5 mM diluted in dimethyl sulfoxide (DMSO) in Hepes buffer with 50 mM CaCl₂ 25 mM pH 8. The trypsin (1mg/ml) was used to determine inhibitory activity. Control experiments, without trypsin, do not detect the enzymatic activity of the samples by itself. The reactions were performed in Hepes buffer, containing 0.25, 0.5 and 0.75mg/ml of extract in the presence of BapNA in a final volume of 1ml. The samples remained at rest in the refrigerator for 1 hour and centrifuged for 15 minutes at 12000 rpm. For all of these samples 50ul of trypsin was then added and incubated at 37°C and absorbance was read at 410nm on a spectrophotometer. The final concentration of 0.75mg/ml for three extracts (propanol, aqueous and ethanol) inhibited trypsin activity by 26%, 15% and 12% respectively. Acknowledgements: CNPq, FAPEMIG, CAPES.