[Objective] The aim was to optimize the fermentation medium and conditions of antibiotic active substances produced by Antarctic psychrotrophic bacterium Rheinheimera sp.97.[Method] Single-factor experiment and orthogonal test were adopted to optimize the fermentation medium of antibiotic active substances produced by Antarctic psychrotrophic bacterium R.sp.97,while the fermentation conditions were optimized by single-factor experiment.[Result] The optimum fermentation medium for the antibiotic active substances production was as follows：tryptone 3.0 g/L,ammonium sulfate 1.0 g/L,starch 2.0 g/L,NaCl 15.0 g/L.The optimized fermentation conditions were as follows：the starting pH of medium was 8.0,fermentation temperature was 10 ℃,liquid volume in Erlenmeyer flask was 30 %（V/V）and inoculation amount was 1%（V/V）.Under the optimized fermentation medium and conditions,the antibacterial activity of R.sp.97 was increased by 18.1%.[Conclusion] This study had provided basis for the antibiotic active substances produced by Antarctic psychrotrophic bacterium R.sp.97.
