Oil is extracted from walnut leaves behind large quantities of defatted press cake that is still rich in valuable nutrients. Aspergillus oryzae and Rhizopus oligosporus, two molds traditionally used in Asia, have the necessary enzymes to use the nutrients in the walnut press cake. Walnuts and the press cake contain ellagitannins, known as precursors for ellagic acid and urolithins. In this study, experiments to optimize the solid-state fermentation of walnut press cake were performed in order to liberate ellagic acid from ellagitannins. Extracts of fermented products were then analyzed with an HPLC-DAD to measure the liberation of ellagic acid from ellagitannins. Good growth of R. oligosporus and A. oryzae mycelia on the walnut press cake was observed. A single mold culture was subjected to a hydration of 0.8 mL/g, an addition of 37.5 mmol/kg acetic acid (AA) and 1% NaCl, and an incubation temperature of 25 °C; these were observed to be good conditions for solid-state fermentation for walnut press cake. The highest ellagic acid concentration was obtained at 48 h. At 72 h, degradation dominated the liberation of ellagic acid.
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