Abstract: The fermentation medium and conditions of Streptomyces canus sp. FIM0916 were optimizied for  producing amphomycin by single-factor and orthogonal test design. The optimized medium formula is 1.5% sucrose, 3.5% soybean meal, 0.1% L-histidine, 0.1% KNO₃ and 0.1% CaCO₃；the optimal fermentation conditions are as follows： filling volume is 120 mL for a 500 mL shake flask, shakers rotating speed is 250 r/min， inoculum size is 2%， temperature 28℃， and cultivation time of seed 54 h; the optimal fermentation time are 5 d. Under optimized conditions, the titer of amphomycin was 248% higher than that of control,  which laid the foundation for subsequent development on amphomycin.

Key words: amphomycin, single factor experiment, orthogonal experiment, fermentation conditions optimization