A strain which can use glucose to produce mannitol was isolated from sugarcane juice and identified as Candida parapsilosis. The production of mannitol under different fermentation conditions was studied in the paper.The recipes of optimal media was determined as 200 g / L glucose,30 g / L yeast extract,0. 05 g / L CaCl2·2H2O,0. 02 g / L FeCl3·2H2O and 0. 5 g / L MgSO4·7H2O based on the single factor test and orthogonal experiments. Under optimal conditions,the production of mannitol was 61. 7 g / L in flask culture. The fermentation enlargement was performed using strain C. parapsilosis in 30-liter fermentor. Mannitol reached the highest production of 80. 3 g / L at72 h according to the curve of fermentation.