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Response Surface Methodology (RSM) was used in the study to optimize the production of vanillin from isoeugenol through fermentation by Aspergillus niger I-1472. Three factors were studied which include amount of isoeugenol, resin (Amberlite XAD-4) and Span 80. During fermentation, isoeugenol as substrate were vortexed with Span 80 and added into the culture on Day 4. Resin (Amberlite XAD-4) was added into the medium the following day. The predicted optimum medium combination consisted of 3.61 g/L of isoeugenol, 5.8% (g/ mL) of Amberlite XAD-4 resin and 0.37% of Span 80 with an expected vanillin production of 0.137 g/L. Verification test showed that the model produced similar predicted and experimental values. [ABSTRACT FROM AUTHOR]