Abstract:[Objective] The objective of this study was to increase the yield of 10-deacetylbaccatin III (10-DAB) and taxol in Cladosporium cladosporioides MD2 through optimizing fermentation conditions. [Methods] The culture conditions of C. cladosporioides MD2 were optimized by single factor experiments to investigate the effect of different initial pH values of medium, culture temperatures, rotational speeds, and culture time on the yield of 10-DAB and taxol. The fermentation medium components were optimized by single factor and orthogonal experiments to investigate the effect of four precursors on the yield of 10-DAB and taxol. [Results] The optimization results were that 1 mL spore suspension of C. cladosporioides MD2 (107?108 spores) was inoculated into 300 mL YES medium with the initial pH of 5.0, which was supplemented with 15 mg/L sodium benzoate, 25 mg/L L-phenylalanine, 5 mg/L serine and 15 mg/L glycine, and then cultured at 28.0 °C and 220 r/min for 12 days. Under the optimized fermentation conditions, the biomass, 10-DAB yield and taxol yield of C. cladosporioides MD2 were respectively 15.5 g/L, 471.5 μg/L and 569.5 μg/L, about 1.3 times, 3.6 times and 3.4 times of that under the initial fermentation conditions. [Conclusion] A suitable fermentation condition of C. cladosporioides MD2 for 10-DAB and taxol production in flask culture was obtained for the first time, and these results provided a reference for its scale-up fermentation.