Response surface methodology (RSM) was successfully applied to enzymatic bio-transformation of 1-naphthol. The experiments were conducted in a closed system containing acetone and sodium acetate buffer, with laccase enzyme. Laccase enzyme used as catalyst was derived from Trametes versicolor (ATCC 200801). The enzymatic bio-transformation rate of 1-naphthol, based on measurements of initial dissolved oxygen (DO) consumption rate in the closed system, was optimized by the application of RSM. The independent variables, which had been found as the most effective variables on the initial DO consumption rate by screening experiments, were determined as medium temperature, pH and acetone content. A quadratic model was developed through RSM in terms of related independent variables to describe the DO consumption rate as the response. Based on contour plots and variance analysis, optimum operational conditions for maximizing initial DO consumption rate, while keeping acetone content at its minimum value, were 301 K of temperature, pH 6 and acetone content of 7% to obtain 9.17 x 10(-3) mM DO/min for initial oxidation rate.