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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 (EC 50 ) 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) (EC 50  = 0.26 ± 0.01 mg/mL) was superior to that of 5-butylmethyl furfural (EC 50  = 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).

Fermentation of Agaricus bisporus for antioxidant activity: response surface optimization, chemical components, and mechanism