Isolation of Anticarcinogenic Isoflavone-conjugated Glycoproteins from a Submerged Liquid Culture of Agaricus blazei Mycelia by the Autolysis Process

Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that iso- flavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anti- carcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soy- bean flakes for 14 days. The liquid culture was autolyzed by incubating at 53℃ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the a utolyzed ABM cultures by 80% etha- nol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-1 80 cell-induced female ICR mouse as- cites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with 1 H-NMR spectral analysis, it con- tained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autol- ysis could be a useful tool to produce antitumor agents.