Optimization of Scale-Up Cultivation Medium for Bacillus subtilis A3M3 in Aquaculture
YE Bo, ZHAO Zhenjun, WANG Xuda, YANG Boxue, LIU Danni, DONG Ying, WANG Xiaoyue, LIU Guilin, ZHANG Qian, LI Shilei
Key Laboratory of Germplasm Improvement and Fine Seed Breeding for Marine Aquatic Animals, LiaoningProvince, Key Laboratory of Protection and Utilization of Aquatic Germplasm Resource, Ministry of Agricultureand Rural Affairs, Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China
Abstract：To enhance the yield of Bacillus subtilis A3M3 under simplified scale-up cultivation conditions, reduce production costs, and promote its application as a water quality regulator and feed additive in aquaculture, the scale-up medium was optimized by Plackett-Burman design and response surface methodology using the viable bacterial density in the broth as the response value. First, the key factors influencing the viable count of B. subtilis A3M3 screened by the Plackett-Burman design were found to be brown sugar, soybean peptone, and KH2PO4. Subsequently, in the steepest ascent experiment, the optimal concentration ranges of these key factors were approached, and thus a three-factor, and three-level Box-Behnken experimental design was implemented for response surface analysis to construct a quadratic polynomial regression model correlating the viabledensity with the three key factors. The data showed that the model fitted well and was used for practical predictions. The optimal proportion of each factor was determined by solving the regression equation, with the final optimized medium formulation as: brown sugar of 14.34 g/L, soybean peptone of 9.93 g/L, KH2PO4 of 1.92 g/L, yeast extract of 2 g/L, MgSO4 of 0.2 g/L, CaCO3 of 0.1 g/L, and MnSO4 of 0.01 g/L. Validation experiments showed that the optimized formulation increased the viabledensity of A3M3 from 2.36×109 cfu/mL before optimization to 3.12×109 cfu/mL, increase by 32.2%. This result was close to the model's predicted value (3.35×109 cfu/mL), significantly enhancing bacterial yield. This is conducive to the large-scale production of B. subtilis A3M3 and widespread application in aquaculture practices.
叶博, 赵振军, 王旭达, 杨博学, 刘丹妮, 董颖, 王笑月, 刘桂林, 张乾, 李石磊. 水产养殖用枯草芽孢杆菌A3M3扩培培养基的优化[J]. 水产科学, 2026, 45(5): 698-708.
YE Bo, ZHAO Zhenjun, WANG Xuda, YANG Boxue, LIU Danni, DONG Ying, WANG Xiaoyue, LIU Guilin, ZHANG Qian, LI Shilei. Optimization of Scale-Up Cultivation Medium for Bacillus subtilis A3M3 in Aquaculture. Fisheries Science, 2026, 45(5): 698-708.
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