Effect of fermentation times and extracting solvents on the in vitro immune potentials of the soluble extracts of mucor-fermented Mao-tofu.

Mucor-fermented Mao-tofu at 3-7 days was extracted using water (pH 4.5 and pH 6.5) and ethanol solutions (40 and 80%, v/v). At protein concentration of 40 μg/mL, all the extracts showed immune activities via activating macrophages, splenocytes, and natural killer cells, which enhances the secretion of interleukin (IL)-2, interferon-γ, tumor necrosis factor-α, IL-1β, IL-6, inducible nitric oxide synthase, and lysozyme, but inhibits the secretion of IL-4. All the extracts exhibited higher activities than that of soybean protein, demonstrating that Mucor-mediated fermentation enhanced the immune potentials of Mao-tofu. Fermentation time of 6 days ensured the extracts the highest activities, whereas lower activities were detected. The highest and lowest activities were detected on using a solution of water (pH 6.5) and 40% ethanol, respectively. The extract obtained using water (pH 6.5) contained more Arg and Cys with immune significance, which partially contributed to its high activities. Both fermentation times and extracting solvents were thus proved to affect the immune activities of soluble Mao-tofu extracts.