The invention discloses a high-yield fermentation strategy for producing androstenedione (ADD) by using recombinant bacillus subtilis. A single factor experiment and an orthogonal experiment show that fermentation medium components, culturing conditions and a full-cell transformation technology of recombinant bacteria bacillus subtilis 168/pMA5-ksdD are optimized, concentration and pH values of maltose, soy peptone to yeast extract (2 to 1), ammonium chloride, potassium chloride, corn steep liquid, cosolvent methyl-beta-cyclodextrin and a substrate androstane (AD) in an optimal medium are determined; and the optimal inoculum size and the culturing conditions are determined. The ADD yield after optimization reaches 1.80g/L which is greatly improved by 177 percent compared with 0.65g/L before optimization. According to the fermentation strategy, based on the research on the fermentation strategy of the recombinant bacteria, the ability of producing ADD by using an AD full-cell transformation technology is greatly improved, and a feasible fermentation strategy is provided for highly yielding ADD in an industrial microorganism one-step fermentation process.