响应面法优化生防暹罗芽孢杆菌JZ1-4-10的发酵参数

蒙耀; 陈红雨; 吴慧芳; 魏琳; 沈硕

doi:10.3969/j.issn.2095-1191.2024.06.012

响应面法优化生防暹罗芽孢杆菌JZ1-4-10的发酵参数

Optimization of fermentation parameters against Bacillus siamensis JZ1-4-10 by response surface method

摘要

摘要: 【目的】优化暹罗芽孢杆菌JZ1-4-10的发酵培养基组分配比和发酵条件，以提高其对马铃薯干腐病菌的抑菌活性，为后续相关生防制剂的工业开发及农业应用打下理论基础。【方法】通过单因素试验筛选菌株JZ1-4-10生长的最佳碳源、氮源、无机盐及适宜浓度，通过响应面法获得菌株JZ1-4-10的最佳发酵培养基组分配比，在此基础上优化菌株JZ1-4-10的发酵温度、pH、接种量、装液量和摇床转速等发酵条件。【结果】菌株JZ1-4-10的最佳碳源、氮源和无机盐分别为玉米粉、大豆蛋白胨和KCl。以玉米粉（A）、大豆蛋白胨（B）和KCl(C)为因变量，菌株JZ1-4-10对马铃薯干腐病病原菌茄病镰刀菌的抑菌活性为响应值，拟合得到菌株JZ1-4-10抑菌率（y）的二次多项回归方程为y=64.02+0.4937A+2.31B-2.19C-1.17AB+138AC-1.81BC-4.73A²-4.58B²-3.86C2，其中，玉米粉、大豆蛋白胨和KCl间的交互作用对菌株JZ1-4-10抑菌活性的影响均达显著水平（P<0.05），尤其玉米粉与KCl及大豆蛋白胨与KCl的交互作用对抑菌活性的影响达极显著水平（P<0.01）。响应面分析优化得到菌株JZ1-4-10的最佳发酵培养基组分配方：玉米粉0.94%、大豆蛋白胨1.23%、KCl 0.72%，且该条件下菌株JZ1-4-10的抑菌活性的实际值为65.20%，与试验预测的64.63%非常接近。经单因素优化后菌株JZ1-4-10的最佳发酵条件为装液量90 mL/250 mL、接种量1.5%、摇床转速160 r/min、培养温度30℃、p H 7。【结论】优化后的培养基提高了菌株JZ1-4-10的产孢量和抑菌活性，优化的发酵方案可用于快速、大批量培养菌株JZ1-4-10菌悬液。

Abstract: 【Objective】 The study aimed to optimize the fermentation culture medium composition formulation and conditions of Bacillus siamensis JZ1-4-10 to improve its antibacterial activity against potato dry rot bacteria, laying a foundation for the subsequent industrial development and agricultural application of related biocontrol agents. 【Method】 The optimal carbon source, nitrogen source, inorganic salt and suitable concentration of B. siamensis JZ1-4-10 were screened by the single factor test. The optimal fermentation culture medium composition formulation of B. siamensis JZ1-4-10 was obtained using response surface methodology. Based on these findings, the fermentation conditions such as fermentation temperature, pH, inoculation amount, liquid volume and rotational speed of strain JZ1-4-10 were optimized. 【Result】 The optimal carbon source, nitrogen source and inorganic salt of B. siamensis JZ1-4-10 were corn flour, soy peptone and KCl respectively. With corn flour（A）, soy peptone（B） and KCl（C） as the dependent variables, and the antibacterial activity of B. siamensis JZ1-4-10 against the potato dry rot pathogen Fusarium solani as the response value, the quadratic multiple regression equation obtained for the inhibition rate（y） of B. siamensis JZ1-4-10 was y=64.02+0.4937A+2.31B-2.19C-1.17AB+138AC-1.81BC-4.73A²-4.58B²-386C2. The interactions among corn flour, soy peptone and KCl significantly influenced the antibacterial activity of B. siamensis JZ1-4-10（P＜0.05）, especially the interactions between corn flour and KCl and between soy peptone and KCl were extremely significant（P＜0.01）. The optimum proportion of fermentation culture medium composition formulation by response surface analysis for B. siamensis JZ1-4-10 were as follows: 0.94% corn flour, 1.23% soy protein and 0.72% KCl. Under this condition, the actual value of antibacterial activity of B. siamensis JZ1-4-10 was 65.20%, which was very close to the predicted value of 64.63%. Similarly, the optimal fermentation conditions optimized by single factors of B. siamensis JZ1-4-10 were liquid volume of 90 mL/250 mL, inoculation amount of 1.5%, rotational speed of 160 r/min, culture temperature of 30 ℃, and pH of 7. 【Conclusion】 The optimized culture medium enhances the sporulation and antibacterial activity of B. siamensis JZ1-4-10, the optimized fermentation method can be used to culture the suspension of strain JZ1-4-10 in fast speed and by large amount.

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