Biotechnology Bulletin ›› 2026, Vol. 42 ›› Issue (4): 332-344.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0996
SU Chang(
), WEN Feng, WANG Ye-lin, SHAO Rui-ying, TANG Jia-jie, XIA Zhan-feng()
Received:2025-09-17
Online:2026-02-09
Published:2026-02-09
Contact:
XIA Zhan-feng
E-mail:1094615208@qq.com;fenge3721@163.com
SU Chang, WEN Feng, WANG Ye-lin, SHAO Rui-ying, TANG Jia-jie, XIA Zhan-feng. Fermentation Optimization of a Surfactin-producing Bacillus Strain and Its Application in Prevention and Control[J]. Biotechnology Bulletin, 2026, 42(4): 332-344.
Fig. 1 Oil ring drainage control group (left) and experimental group (right)
编码水平 Coding proficiency | A 麦芽糖Maltose （g/L） | B 酵母浸粉Yeast （g/L） | C 氯化钠NaCl （g/L） | D 温度Temperature （ ℃） | E 发酵时间 Fermented time （h） | F pH | G 转速 Rotational speed （r/min） | H 装液量 Loaded liquid （mL） |
|---|---|---|---|---|---|---|---|---|
| 低Low（-1） | 8 | 2 | 4 | 25 | 48 | 5 | 120 | 100 |
| 高Hight（1） | 16 | 8 | 10 | 35 | 96 | 8 | 160 | 200 |
Table 1 Types and levels of factors in the Plackett-Burman experimental design
编码水平 Coding proficiency | A 麦芽糖Maltose （g/L） | B 酵母浸粉Yeast （g/L） | C 氯化钠NaCl （g/L） | D 温度Temperature （ ℃） | E 发酵时间 Fermented time （h） | F pH | G 转速 Rotational speed （r/min） | H 装液量 Loaded liquid （mL） |
|---|---|---|---|---|---|---|---|---|
| 低Low（-1） | 8 | 2 | 4 | 25 | 48 | 5 | 120 | 100 |
| 高Hight（1） | 16 | 8 | 10 | 35 | 96 | 8 | 160 | 200 |
因素 Factor | 编码水平 Horizontal encoding | |||
|---|---|---|---|---|
| -1 | 0 | 1 | ||
| 酵母浸粉 Yeast （g/L） | 6 | 7 | 8 | |
| 麦芽糖 Maltose （g/L） | 10.5 | 9 | 7.5 | |
| 摇床转速 Rotational speed （r/min） | 150 | 160 | 170 | |
Table 2 Variables and their levels in the Box-Behnken experiment
因素 Factor | 编码水平 Horizontal encoding | |||
|---|---|---|---|---|
| -1 | 0 | 1 | ||
| 酵母浸粉 Yeast （g/L） | 6 | 7 | 8 | |
| 麦芽糖 Maltose （g/L） | 10.5 | 9 | 7.5 | |
| 摇床转速 Rotational speed （r/min） | 150 | 160 | 170 | |
Fig. 2 Surfactin standard curve
Fig. 3 Optimization of the fermentation medium for strain TRM82479Different lowercase letters indicate significant difference at 0.05 level, the same below
组别 Group | 碳源种类 Carbon source （g/L） | 氮源种类 Nitrogen source （g/L） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|
| 1 | 淀粉 | 大豆蛋白胨 | 20.15 |
| 2 | 燕麦 | 酸水解酪蛋白 | 40.44 |
| 3 | 燕麦 | 大豆蛋白胨 | 18.13 |
| 4 | 麦芽糖 | 酸水解酪蛋白 | 29.83 |
| 5 | 燕麦 | 酵母浸粉 | 38.24 |
| 6 | 淀粉 | 酵母浸粉 | 30.78 |
| 7 | 麦芽糖 | 酵母浸粉 | 51.03 |
| 8 | 淀粉 | 酸水解酪蛋白 | 30.98 |
| 9 | 麦芽糖 | 大豆蛋白胨 | 39.11 |
Table 3 Combined table and results of the orthogonal experimental group of carbon and nitrogen sources
组别 Group | 碳源种类 Carbon source （g/L） | 氮源种类 Nitrogen source （g/L） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|
| 1 | 淀粉 | 大豆蛋白胨 | 20.15 |
| 2 | 燕麦 | 酸水解酪蛋白 | 40.44 |
| 3 | 燕麦 | 大豆蛋白胨 | 18.13 |
| 4 | 麦芽糖 | 酸水解酪蛋白 | 29.83 |
| 5 | 燕麦 | 酵母浸粉 | 38.24 |
| 6 | 淀粉 | 酵母浸粉 | 30.78 |
| 7 | 麦芽糖 | 酵母浸粉 | 51.03 |
| 8 | 淀粉 | 酸水解酪蛋白 | 30.98 |
| 9 | 麦芽糖 | 大豆蛋白胨 | 39.11 |
Fig. 4 Optimization of fermentation conditions for strain TRM82479
组别 Group | Plackett-Burman实验因素 Plackett-Burman experimental factors | 排油圈直径 Diameter of oil ring drainage （mm） | |||||||
|---|---|---|---|---|---|---|---|---|---|
| A | B | C | D | E | F | G | H | ||
| 1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | 32.87 |
| 2 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | 71.88 |
| 3 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | 27.79 |
| 4 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 66.40 |
| 5 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 36.01 |
| 6 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 31.76 |
| 7 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | 37.65 |
| 8 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 41.94 |
| 9 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 65.53 |
| 10 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | 60.86 |
| 11 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 22.83 |
| 12 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | 18.11 |
Table 4 Plackett-Burman experimental design and results
组别 Group | Plackett-Burman实验因素 Plackett-Burman experimental factors | 排油圈直径 Diameter of oil ring drainage （mm） | |||||||
|---|---|---|---|---|---|---|---|---|---|
| A | B | C | D | E | F | G | H | ||
| 1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | 32.87 |
| 2 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | 71.88 |
| 3 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | 27.79 |
| 4 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 66.40 |
| 5 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 36.01 |
| 6 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 31.76 |
| 7 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | 37.65 |
| 8 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 41.94 |
| 9 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 65.53 |
| 10 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | 60.86 |
| 11 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 22.83 |
| 12 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | 18.11 |
来源 Source | 平方和 Sum of squares | 自由度 DF | 均方 Mean square | F值 F-value | P值 P-value | 显著性 Significance |
|---|---|---|---|---|---|---|
| 模型 Model | 3 699.19 | 8 | 462.40 | 18.73 | 0.017 4 | * |
| A | 265.27 | 1 | 265.27 | 10.74 | 0.046 5 | * |
| B | 2 278.11 | 1 | 2 278.11 | 92.25 | 0.002 4 | ** |
| C | 262.95 | 1 | 262.95 | 10.65 | 0.047 | * |
| D | 68.23 | 1 | 68.23 | 2.76 | 0.195 1 | |
| E | 39.07 | 1 | 39.07 | 1.58 | 0.297 4 | |
| F | 70.7 | 1 | 70.7 | 2.86 | 0.189 2 | |
| G | 651.02 | 1 | 651.02 | 26.36 | 0.014 3 | * |
| H | 63.85 | 1 | 63.85 | 2.59 | 0.206 2 | |
| 残差 Residual | 74.08 | 3 | 24.69 | |||
| 总和 Cor total | 3 773.27 | 11 | ||||
| R2 | 0.980 4 | Predicted R2 | 0.685 9 | |||
| R2adj | 0.928 0 | Adeq precision | 11.991 4 |
Table 5 Plackett-Burman experiment factors, levels and significance analysis
来源 Source | 平方和 Sum of squares | 自由度 DF | 均方 Mean square | F值 F-value | P值 P-value | 显著性 Significance |
|---|---|---|---|---|---|---|
| 模型 Model | 3 699.19 | 8 | 462.40 | 18.73 | 0.017 4 | * |
| A | 265.27 | 1 | 265.27 | 10.74 | 0.046 5 | * |
| B | 2 278.11 | 1 | 2 278.11 | 92.25 | 0.002 4 | ** |
| C | 262.95 | 1 | 262.95 | 10.65 | 0.047 | * |
| D | 68.23 | 1 | 68.23 | 2.76 | 0.195 1 | |
| E | 39.07 | 1 | 39.07 | 1.58 | 0.297 4 | |
| F | 70.7 | 1 | 70.7 | 2.86 | 0.189 2 | |
| G | 651.02 | 1 | 651.02 | 26.36 | 0.014 3 | * |
| H | 63.85 | 1 | 63.85 | 2.59 | 0.206 2 | |
| 残差 Residual | 74.08 | 3 | 24.69 | |||
| 总和 Cor total | 3 773.27 | 11 | ||||
| R2 | 0.980 4 | Predicted R2 | 0.685 9 | |||
| R2adj | 0.928 0 | Adeq precision | 11.991 4 |
组别 Group | 步长 Step size | 酵母浸粉 Yeast （g/L） | 麦芽糖 Maltose （g/L） | 转速 Rotational speed （r/min） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|---|---|
| 1 | 0 | 5 | 12 | 140 | 58.73b |
| 2 | 0+1Δ | 6 | 10.5 | 150 | 61.56b |
| 3 | 0+2Δ | 7 | 9 | 160 | 70.55a |
| 4 | 0+3Δ | 8 | 7.5 | 170 | 60.94b |
| 5 | 0+4Δ | 9 | 6 | 180 | 43.48c |
| 6 | 0+5Δ | 10 | 4.5 | 190 | 39.05c |
Table 6 Design of the steep slope climbing experiment
组别 Group | 步长 Step size | 酵母浸粉 Yeast （g/L） | 麦芽糖 Maltose （g/L） | 转速 Rotational speed （r/min） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|---|---|
| 1 | 0 | 5 | 12 | 140 | 58.73b |
| 2 | 0+1Δ | 6 | 10.5 | 150 | 61.56b |
| 3 | 0+2Δ | 7 | 9 | 160 | 70.55a |
| 4 | 0+3Δ | 8 | 7.5 | 170 | 60.94b |
| 5 | 0+4Δ | 9 | 6 | 180 | 43.48c |
| 6 | 0+5Δ | 10 | 4.5 | 190 | 39.05c |
组别 Group | 酵母浸粉 Yeast （g/L） | 麦芽糖 Maltose （g/L） | 转速 Rotational speed（r/min） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|---|
| 1 | -1 | -1 | 0 | 54.81 |
| 2 | 1 | -1 | 0 | 58.79 |
| 3 | -1 | 1 | 0 | 48.86 |
| 4 | 1 | 1 | 0 | 58.38 |
| 5 | -1 | 0 | -1 | 44.28 |
| 6 | 1 | 0 | -1 | 52.73 |
| 7 | -1 | 0 | 1 | 49.84 |
| 8 | 1 | 0 | 1 | 55.35 |
| 9 | 0 | -1 | -1 | 52.81 |
| 10 | 0 | 1 | -1 | 53.01 |
| 11 | 0 | -1 | 1 | 57.97 |
| 12 | 0 | 1 | 1 | 55.53 |
| 13 | 0 | 0 | 0 | 74.84 |
| 14 | 0 | 0 | 0 | 76.73 |
| 15 | 0 | 0 | 0 | 73.84 |
| 16 | 0 | 0 | 0 | 75.84 |
| 17 | 0 | 0 | 0 | 75.68 |
Table 7 Box-Behnken experimental design and results
组别 Group | 酵母浸粉 Yeast （g/L） | 麦芽糖 Maltose （g/L） | 转速 Rotational speed（r/min） | 排油圈直径 Diameter of oil ring drainage （mm） |
|---|---|---|---|---|
| 1 | -1 | -1 | 0 | 54.81 |
| 2 | 1 | -1 | 0 | 58.79 |
| 3 | -1 | 1 | 0 | 48.86 |
| 4 | 1 | 1 | 0 | 58.38 |
| 5 | -1 | 0 | -1 | 44.28 |
| 6 | 1 | 0 | -1 | 52.73 |
| 7 | -1 | 0 | 1 | 49.84 |
| 8 | 1 | 0 | 1 | 55.35 |
| 9 | 0 | -1 | -1 | 52.81 |
| 10 | 0 | 1 | -1 | 53.01 |
| 11 | 0 | -1 | 1 | 57.97 |
| 12 | 0 | 1 | 1 | 55.53 |
| 13 | 0 | 0 | 0 | 74.84 |
| 14 | 0 | 0 | 0 | 76.73 |
| 15 | 0 | 0 | 0 | 73.84 |
| 16 | 0 | 0 | 0 | 75.84 |
| 17 | 0 | 0 | 0 | 75.68 |
来源 Source | 平方和 Sum of squares | 自由度 DF | 均方 Mean square | F值 F-value | P值 P-value | 显著性 Significance |
|---|---|---|---|---|---|---|
| 模型 Model | 1 885.85 | 9 | 209.54 | 209.76 | <0.000 1 | ** |
| A | 94.23 | 1 | 94.23 | 94.33 | <0.000 1 | ** |
| B | 9.23 | 1 | 9.23 | 9.24 | 0.018 9 | * |
| C | 31.46 | 1 | 31.46 | 31.49 | 0.000 8 | ** |
| AB | 7.68 | 1 | 7.68 | 7.69 | 0.027 6 | * |
| AC | 2.17 | 1 | 2.17 | 2.17 | 0.183 9 | |
| BC | 1.75 | 1 | 1.75 | 1.75 | 0.227 6 | |
| A² | 629.49 | 1 | 629.49 | 630.15 | <0.000 1 | ** |
| B² | 265.98 | 1 | 265.98 | 266.26 | <0.000 1 | ** |
| C² | 669.22 | 1 | 669.22 | 669.92 | <0.000 1 | ** |
| 残差 Residual | 6.99 | 7 | 0.999 | |||
| 失拟 Lack of fit | 2.18 | 3 | 0.726 5 | 0.603 8 | 0.646 3 | |
| 变异系数CV（%） | 1.67% | 校正决定系数R2 | 0.996 3 | |||
| 复相关系数R2adj | 0.991 6 | |||||
| 预测系数 Pred R2 | 0.977 6 | |||||
| 精密度 Adeq precision | 40.421 9 |
Table 8 ANOVA of the quadratic model based on Box-Behnken design
来源 Source | 平方和 Sum of squares | 自由度 DF | 均方 Mean square | F值 F-value | P值 P-value | 显著性 Significance |
|---|---|---|---|---|---|---|
| 模型 Model | 1 885.85 | 9 | 209.54 | 209.76 | <0.000 1 | ** |
| A | 94.23 | 1 | 94.23 | 94.33 | <0.000 1 | ** |
| B | 9.23 | 1 | 9.23 | 9.24 | 0.018 9 | * |
| C | 31.46 | 1 | 31.46 | 31.49 | 0.000 8 | ** |
| AB | 7.68 | 1 | 7.68 | 7.69 | 0.027 6 | * |
| AC | 2.17 | 1 | 2.17 | 2.17 | 0.183 9 | |
| BC | 1.75 | 1 | 1.75 | 1.75 | 0.227 6 | |
| A² | 629.49 | 1 | 629.49 | 630.15 | <0.000 1 | ** |
| B² | 265.98 | 1 | 265.98 | 266.26 | <0.000 1 | ** |
| C² | 669.22 | 1 | 669.22 | 669.92 | <0.000 1 | ** |
| 残差 Residual | 6.99 | 7 | 0.999 | |||
| 失拟 Lack of fit | 2.18 | 3 | 0.726 5 | 0.603 8 | 0.646 3 | |
| 变异系数CV（%） | 1.67% | 校正决定系数R2 | 0.996 3 | |||
| 复相关系数R2adj | 0.991 6 | |||||
| 预测系数 Pred R2 | 0.977 6 | |||||
| 精密度 Adeq precision | 40.421 9 |
Fig. 5 Response surfaces and contour lines of the interactions between various factors on the diameter of the oil discharge ring
供试虫源 Insect source tested | 虫态 Instar | 死亡率 Mortality （%） | 校正死亡率 Corrected mortality （%） |
|---|---|---|---|
| 棉铃虫 Helicoverpa armigera | 2龄幼虫 2nd instar larva | （23.33±7.64）e | （21.05±0）e |
| 草地贪夜蛾 Spodoptera frugiperda | 2龄幼虫 2nd instar larva | （36.67±2.89）d | （32.15±0.99）d |
| 甜菜夜蛾 Spodoptera exigua | 2龄幼虫 2nd instar larva | （65±5.00）b | （61.11±5.56）b |
| 玉米螟 Ostrinia nubilalis | 2龄幼虫 2nd instar larva | （35±5.00）d | （29.13±3.68）de |
| 黏虫 Mythimna separata | 2龄幼虫 2nd instar larva | （51.64±7.64）c | （48.15±8.52）c |
| 斜纹夜蛾 Prodenia litura | 2龄幼虫 2nd instar larva | （66.67±2.89）b | （64.23±3.77）b |
| 蚜虫 Aphis gossypii | 若虫期 Nymphal stage | （91.67±2.89）a | （91.23±3.04）a |
Table 9 Insecticidal activity of fermentation broth of TRM82479
供试虫源 Insect source tested | 虫态 Instar | 死亡率 Mortality （%） | 校正死亡率 Corrected mortality （%） |
|---|---|---|---|
| 棉铃虫 Helicoverpa armigera | 2龄幼虫 2nd instar larva | （23.33±7.64）e | （21.05±0）e |
| 草地贪夜蛾 Spodoptera frugiperda | 2龄幼虫 2nd instar larva | （36.67±2.89）d | （32.15±0.99）d |
| 甜菜夜蛾 Spodoptera exigua | 2龄幼虫 2nd instar larva | （65±5.00）b | （61.11±5.56）b |
| 玉米螟 Ostrinia nubilalis | 2龄幼虫 2nd instar larva | （35±5.00）d | （29.13±3.68）de |
| 黏虫 Mythimna separata | 2龄幼虫 2nd instar larva | （51.64±7.64）c | （48.15±8.52）c |
| 斜纹夜蛾 Prodenia litura | 2龄幼虫 2nd instar larva | （66.67±2.89）b | （64.23±3.77）b |
| 蚜虫 Aphis gossypii | 若虫期 Nymphal stage | （91.67±2.89）a | （91.23±3.04）a |
药剂 Insecticides | 虫口基数Number | 药后1 d One day later | 药后2 d Two days later | 药后7 d Seven days later | |||
|---|---|---|---|---|---|---|---|
残虫数 Number | 校正防效 Control efficacy （%） | 残虫数 Number | 校正防效 Control efficacy （%） | 残虫数 Number | 校正防效 Control efficacy （%） | ||
| 108 CFU /mL菌液 | 86 | 19 | 77.61±6.43a | 14 | 80.45±7.98a | 16 | 77.33±6.79a |
| 107 CFU /mL菌液 | 74 | 23 | 73.82±5.53ab | 20 | 72.05±8.80ab | 18 | 70.48±4.72ab |
| 106 CFU /mL菌液 | 75 | 30 | 70.78±6.79ab | 23 | 71.29±7.27ab | 22 | 66.26±2.38b |
| 25%氟啶虫酰胺 | 97 | 27 | 67.03±3.59b | 24 | 68.10±2.21b | 36 | 56.95±9.02c |
| 清水 | 71 | 66 | _ | 63 | _ | 60 | _ |
Table 10 Field efficacy of TRM82479 fermentation broth against cotton aphids
药剂 Insecticides | 虫口基数Number | 药后1 d One day later | 药后2 d Two days later | 药后7 d Seven days later | |||
|---|---|---|---|---|---|---|---|
残虫数 Number | 校正防效 Control efficacy （%） | 残虫数 Number | 校正防效 Control efficacy （%） | 残虫数 Number | 校正防效 Control efficacy （%） | ||
| 108 CFU /mL菌液 | 86 | 19 | 77.61±6.43a | 14 | 80.45±7.98a | 16 | 77.33±6.79a |
| 107 CFU /mL菌液 | 74 | 23 | 73.82±5.53ab | 20 | 72.05±8.80ab | 18 | 70.48±4.72ab |
| 106 CFU /mL菌液 | 75 | 30 | 70.78±6.79ab | 23 | 71.29±7.27ab | 22 | 66.26±2.38b |
| 25%氟啶虫酰胺 | 97 | 27 | 67.03±3.59b | 24 | 68.10±2.21b | 36 | 56.95±9.02c |
| 清水 | 71 | 66 | _ | 63 | _ | 60 | _ |
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