OBJECTIVE To improve the yield and quality of L-lactic acid by Rhizopus oryzae, we aim to understand the relationship between inorganic salts utilization and the L-lactic acid metabolism of the strain RLC41-6, through systematic analysis of the effects of zinc ion concentration on the production of L-lactic acid and the Lactic Dehydrogenase (LDH) activity. METHODS Rhizopus oryzae was cultured at 36 degrees C for 36h with different quantity of ZnSO4 in fermentation medium. The fermentation products were monitored by reversed-phase high performance liquid chromatography (RP-HPLC), LDH isoenzyme composition in the cell was analysed by non-denatured polyacrylamide gel electrophoresis (PAGE). RESULTS Our results showed that the concentration of ZnSO4 in medium could modulate the expression of LDH isoenzyme except LDH1, especially stimulated the expression of LDH4 and LDH5. When initial concentration of ZnSO4 is above 0.02%, the LDH4 and LDH5 reached the highest level. However, the activity of LDH was inhibited by higher concentration zinc ion in extracellular environment. When ZnSO4 concentration is 0.02%, LDH activity reaches its maximum 200U/mL, the HPLC assay showed only L-lactic in the fermentation products (137 g/L), while the conversion rate of glucose to lactic acid is 91%. CONCLUSIONS Zinc ion can regulate the metabolic processes of Rhizopus oryzae and modulate the types of the final fermentation products. An optimal concentration of ZnSO4 can not only facilitate the LDH expression but also prevent pyruvate from transformation into the malic acid and fumaric acid during the metabolism process, thereby enhance the metabolism of glucose to lactic acid of Rhizopus oryzae.