The invention belongs to the field of fermentation engineering, and particularly relates to a method for improving yield of bacitracin by microbial fermentation. According to the method, by a step 1) of optimization of organic compounding of a nitrogen source and fermentation conditions, stability of fermentation units between batches is ensured, and the content of impurities F is reduced; and by a step 2) of adding superparamagnetic microspheres during the synthesis of bacitracin, feedback inhibition of produced bacteria by fermentation products is reduced. After fermentation, the fermentation unit of bacitracin can reach 1228U/ml, which can increase the yield of bacitracin by 123%, improve the productivity and reduce energy consumption and waste discharge.