Objective This study aimed to screen for a high-mupirocin-producing strain and improve the fermentation level.Methods The breeding technology of atmospheric room temperature plasma mutation(ARTP)was used to screen the strain PF 16002059,which produced mupirocin.At the same time,the resistance to self-produced metabolites was used as the selection pressure,and the mutants were screened by shake flask fermentation.Results A stable and high-yield production strain PF22033156 was obtained.The shake flask fermentation level of the strain PF22033156 reached 7062 mg/L,which was 98%higher than the original strain PF 16002059.Conclusion ARTP mutagenesis combined with resistance to self-produced metabolites were used to screen for the new strain PF22033156(Pseudomonas fluorescens PF22033156)in this study,which significantly increased the production of mupirocinm,and the genetic character of the mutant strain was stable,which made it suitable for industrial production.