Fermentation ofAgaricus bisporusfor antioxidant activity: response surface optimization, chemical components, and mechanism

Agaricus bisporus is one of the most widely cultivated edible mushrooms in the world. The chemical components of A. bisporus have a wide range of biological activities. In order to deeply understand the antioxidant properties of A. bisporus, this study conducted an investigation on the components of A. bisporus fermentation. Through the single factor experiment and response surface optimization, it was found that when the C/N ratio was 45:1, the inoculum concentration was 10%, and the fermentation time was 7 d, the n-butanol extract of the fermentation product had the strongest scavenging capacity for free radical generated through 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulphonic acid (ABTS·+). The concentration for 50% of the maximal effect (EC50) was 0.33 ± 0.01 mg/mL. Moreover, in order to identify the two main components, the elution-extrusion counter-current chromatography (EECCC) was employed for separation, where 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) and 5-(butoxymethyl) furfural were obtained. The antioxidant activity of 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) (EC50 = 0.26 ± 0.01 mg/mL) was superior to that of 5-butylmethyl furfural (EC50 = 1.52 ± 0.02 mg/mL), indicating that 5,5'-oxy-dimethyl-bis(2-furfuraldehyde) was the main antioxidant in the fermentation products. The thermodynamic parameters and frontier molecular orbitals of 5,5'-oxy-dimethyl-bis (2-furanaldehyde) was evaluated by density functional theory (DFT). The result indicated 5,5'-oxy-dimethyl-bis(2-furanaldehyde) scavenged free radicals in polar media through single electron transfer followed by proton transfer (SET-PT).