Kojic acid production from cocoa juice by Aspergillus flavus entrapped in calcium alginate

Sixteen microorganisms of Aspergillus strains were screened for production of kojic acid using cocoa juice as carbon source. Only Aspergillus flavus ATCC 9179 was found to produce the acid in low yield (22 mg/ml). Calcium alginate immobilization of the cells was used under optimum conditions to maximize the yield of kojic acid (60 mg/ml). Cultures were incubated in the medium with 50% of cocoa juice added in pulses of 8 ml each every 96 hours, and 4% methanol, pH 3.5, 150 rpm, 26 degrees C for three weeks. The incubations were monitored by thin layer and high pressure liquid chromatography. Kojic acid was extracted from the culture broth by organic solvent, concentrated and crystallized. The chemical identity of kojic acid was determined by HPLC, MS, 1H- and 13C-NMR spectroscopy.