The cell growth and total accumulation of bioactive metabolites were significantly improved by Cu2+ addition during the submerged fermentation of medicinal mushroom Ganoderma lucidum. A mathematical model, constructed by response surface methodology combination with full factorial design, was applied to study the synergic effect of Cu2+ addition concentration and addition time. The optimal Cu2+ inducement strategy for the cell growth were different from those for the biosynthesis of ganoderic acid (GA) and Ganoderma polysaccharide. A multiple additions strategy of Cu2+ by adding each 1 mM Cu2+ on day 2, 6, 8 and 2 mM Cu2+ on day 4 was developed to enhance total accumulation of GA and extracellular polysaccharides. The highest GA content reached 3.0 +/- 0.1 mg per 100 mg DW, which was increased by 76.5% and 33.9% compared with the control without Cu2+ addition and the peak value predicted by the constructed mathematical model, respectively. While, relatively higher addition concentration of Cu2+ (i.e., 5 mM) on the culture of day 4 led to higher content and total production of intracellular polysaccharides. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 26: 417-423, 2010