A multi-step response surface methodology was successfully applied to optimize the biosorption conditions for the maximum removal of Cu(II) ions from aqueous solutions using Trametes versicolor fungi as a biosorbent. In the first step, the most effective medium factors, which are pH, temperature and initial Cu(II) concentration, on biosorption of Cu(II), were determined through Plackett-Burman Design. Then steepest accent followed by central composite design steps were utilized to evaluate the optimum biosorption conditions for the maximum Cu(II) ions removal. Based on the statistic analysis; the optimum conditions were obtained 5.51, 20.13 degrees C and 60.98 mg/L as medium pH, medium temperature and initial Cu(II) concentration, respectively. Finally the analysis of variance (ANOVA) of central composite design showed the proposed quadratic model fitted experimental data very well. (C) 2010 Elsevier Ltd. All rights reserved.