Journal of Agricultural Science and Technology ›› 2022, Vol. 24 ›› Issue (8): 133-143.DOI: 10.13304/j.nykjdb.2022.0422
• ANIMAL AND PLANT HEALTH • Previous Articles
Jinrong LI1(), Tong ZHOU2, Yiqi LIN1, Zuohua HUANG1, Liangjing SHENG1, Feiping ZHANG1, Songqing WU3(
)
Received:
2022-05-19
Accepted:
2022-06-20
Online:
2022-08-15
Published:
2022-08-22
Contact:
Songqing WU
李金荣1(), 周彤2, 林一琪1, 黄祚骅1, 盛亮菁1, 张飞萍1, 吴松青3(
)
通讯作者:
吴松青
作者简介:
李金荣 E-mail:jinrongLi@fafu.edu.cn;
基金资助:
CLC Number:
Jinrong LI, Tong ZHOU, Yiqi LIN, Zuohua HUANG, Liangjing SHENG, Feiping ZHANG, Songqing WU. Optimization of Serratia marcescens BRC-CXG2 Fermentation Medium Based on Response Surface Methodology[J]. Journal of Agricultural Science and Technology, 2022, 24(8): 133-143.
李金荣, 周彤, 林一琪, 黄祚骅, 盛亮菁, 张飞萍, 吴松青. 基于响应面法的粘质沙雷氏菌BRC-CXG2发酵培养基优化[J]. 中国农业科技导报, 2022, 24(8): 133-143.
因素 Factor | 水平 Level | |
---|---|---|
低 Low(-1) | 高 High(+1) | |
X1∶油酸 Oleic acid/(g·L-1) | 10 | 15 |
X2∶葡萄糖 Glucose/(g·L-1) | 10 | 12 |
X3∶虚拟项 Virtual items | / | / |
X4∶蚕蛹粉 Soybean cake meal/(g·L-1) | 8 | 12 |
X5∶酵母粉 Yeast extract/(g·L-1) | 8 | 12 |
X6∶虚拟项 Virtual items | / | / |
X7∶硫酸镁 MgSO4/(g·L-1) | 1 | 2 |
X8∶磷酸氢二钠 Na2HPO4/(g·L-1) | 2 | 3 |
X9∶虚拟项 Virtual items | / | / |
X10∶接种量 Inoculatin amount/ % | 1 | 2 |
X11∶装液量 Liquid volume /(mL/250 mL) | 50 | 100 |
Table 1 Factors and levels of Plackett-Burman test
因素 Factor | 水平 Level | |
---|---|---|
低 Low(-1) | 高 High(+1) | |
X1∶油酸 Oleic acid/(g·L-1) | 10 | 15 |
X2∶葡萄糖 Glucose/(g·L-1) | 10 | 12 |
X3∶虚拟项 Virtual items | / | / |
X4∶蚕蛹粉 Soybean cake meal/(g·L-1) | 8 | 12 |
X5∶酵母粉 Yeast extract/(g·L-1) | 8 | 12 |
X6∶虚拟项 Virtual items | / | / |
X7∶硫酸镁 MgSO4/(g·L-1) | 1 | 2 |
X8∶磷酸氢二钠 Na2HPO4/(g·L-1) | 2 | 3 |
X9∶虚拟项 Virtual items | / | / |
X10∶接种量 Inoculatin amount/ % | 1 | 2 |
X11∶装液量 Liquid volume /(mL/250 mL) | 50 | 100 |
Fig. 1 Colonies number of Serratia marcescens under different temperatures, loading volumes and rotational speedsNote: Different lowercase letters in the figure indicate significant differences at P<0.05 level.
Fig. 2 Colonies number of Serratia marcescens under different carbon sourcesNote: Different lowercase letters in the figure indicate significant differences at P<0.05 level.
Fig. 3 Colonies number of Serratia marcescens under different nitrogen sourcesNote: Different lowercase letters in the figure indicate significant differences at P<0.05 level.
Fig. 4 Colonies number of Serratia marcescens under different inorganic salt sourcesNote: Different lowercase letters in the figure indicate significant differences at P<0.05 level.
编号 Number | X1 | X2 | X3 | X4 | X5 | X6 | X7 | X8 | X9 | X10 | X11 | 菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 15.133 |
2 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | 7.022 |
3 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 3.980 |
4 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | 13.367 |
5 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | 1.633 |
6 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | 16.900 |
7 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1.637 |
8 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 4.533 |
9 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 10.967 |
10 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | 18.533 |
11 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 6.333 |
12 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | 3.100 |
Table 2 Design and results of Plackett-Burman test
编号 Number | X1 | X2 | X3 | X4 | X5 | X6 | X7 | X8 | X9 | X10 | X11 | 菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 15.133 |
2 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | 7.022 |
3 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 3.980 |
4 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | 13.367 |
5 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | -1 | 1.633 |
6 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1 | 16.900 |
7 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 1 | 1.637 |
8 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 1 | 4.533 |
9 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | -1 | 10.967 |
10 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 1 | 18.533 |
11 | 1 | -1 | 1 | 1 | 1 | -1 | -1 | -1 | 1 | -1 | 1 | 6.333 |
12 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | -1 | 3.100 |
方差来源 Source | 平方和 Sum of squares | 自由度 Degree of freedom | 均方 Mean square | F值 F value | P值 P value |
---|---|---|---|---|---|
模型 Model | 40 471.24 | 8 | 5 058.91 | 22.87 | 0.013 1 |
油酸 Oleic acid | 2 691.61 | 1 | 2 691.61 | 12.17 | 0.039 8 |
葡萄糖 Glucose | 10 783.21 | 1 | 10 783.21 | 48.75 | 0.006 0 |
蚕蛹粉 Soybean cake meal | 17 414.54 | 1 | 17 414.54 | 77.50 | 0.003 1 |
酵母粉 Yeast extract | 1 384.17 | 1 | 1 384.17 | 6.26 | 0.087 6 |
硫酸镁 MgSO4 | 176.95 | 1 | 176.95 | 0.80 | 0.437 0 |
磷酸氢二钠 Na2HPO4 | 2 072.97 | 1 | 2 072.97 | 9.37 | 0.054 9 |
接种量 Inoculatin amount | 5 837.96 | 1 | 5 837.96 | 26.39 | 0.014 3 |
装液量 Liquid volume | 382.84 | 1 | 382.84 | 1.73 | 0.279 8 |
残差Residual | 663.55 | 3 | 221.18 | — | — |
总和Cor Total | 41 134.80 | 11 | — | — | — |
Table 3 Analysis of variance for the Plackett-Burman test
方差来源 Source | 平方和 Sum of squares | 自由度 Degree of freedom | 均方 Mean square | F值 F value | P值 P value |
---|---|---|---|---|---|
模型 Model | 40 471.24 | 8 | 5 058.91 | 22.87 | 0.013 1 |
油酸 Oleic acid | 2 691.61 | 1 | 2 691.61 | 12.17 | 0.039 8 |
葡萄糖 Glucose | 10 783.21 | 1 | 10 783.21 | 48.75 | 0.006 0 |
蚕蛹粉 Soybean cake meal | 17 414.54 | 1 | 17 414.54 | 77.50 | 0.003 1 |
酵母粉 Yeast extract | 1 384.17 | 1 | 1 384.17 | 6.26 | 0.087 6 |
硫酸镁 MgSO4 | 176.95 | 1 | 176.95 | 0.80 | 0.437 0 |
磷酸氢二钠 Na2HPO4 | 2 072.97 | 1 | 2 072.97 | 9.37 | 0.054 9 |
接种量 Inoculatin amount | 5 837.96 | 1 | 5 837.96 | 26.39 | 0.014 3 |
装液量 Liquid volume | 382.84 | 1 | 382.84 | 1.73 | 0.279 8 |
残差Residual | 663.55 | 3 | 221.18 | — | — |
总和Cor Total | 41 134.80 | 11 | — | — | — |
编号 Number | 葡萄糖 Glucose/(g·L-1) | 蚕蛹粉 Soybean cake meal/(g·L-1) | 接种量 Inoculatin amount/% | 菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|
1 | 16 | 20 | 3.0 | 13.73 |
2 | 18 | 22 | 3.5 | 21.00 |
3 | 20 | 24 | 4.0 | 31.77 |
4 | 22 | 26 | 4.5 | 34.37 |
5 | 24 | 28 | 5.0 | 20.93 |
6 | 26 | 30 | 5.5 | 13.13 |
Table 4 Design and results from the steepest ascent experiment
编号 Number | 葡萄糖 Glucose/(g·L-1) | 蚕蛹粉 Soybean cake meal/(g·L-1) | 接种量 Inoculatin amount/% | 菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|
1 | 16 | 20 | 3.0 | 13.73 |
2 | 18 | 22 | 3.5 | 21.00 |
3 | 20 | 24 | 4.0 | 31.77 |
4 | 22 | 26 | 4.5 | 34.37 |
5 | 24 | 28 | 5.0 | 20.93 |
6 | 26 | 30 | 5.5 | 13.13 |
编号 Number | 葡萄糖 Glucose/(g·L-1) | 蚕蛹粉 Soybean cake meal/(g·L-1) | 接种量 Inoculatin amount/% | Y:菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|
1 | 15 | 20 | 5 | 10.5 |
2 | 25 | 20 | 5 | 49.2 |
3 | 15 | 30 | 5 | 24.9 |
4 | 25 | 30 | 5 | 29.7 |
5 | 15 | 25 | 2.5 | 3.0 |
6 | 25 | 25 | 2.5 | 32.0 |
7 | 15 | 25 | 7.5 | 8.1 |
8 | 25 | 25 | 7.5 | 34.7 |
9 | 20 | 20 | 2.5 | 37.8 |
10 | 20 | 30 | 2.5 | 5.4 |
11 | 20 | 20 | 7.5 | 20.0 |
12 | 20 | 30 | 7.5 | 26.5 |
13 | 20 | 25 | 5 | 44.3 |
14 | 20 | 25 | 5 | 47.6 |
15 | 20 | 25 | 5 | 50.4 |
16 | 20 | 25 | 5 | 59.5 |
17 | 20 | 25 | 5 | 55.5 |
Table 5 Experimental design and results of response surface analysis method
编号 Number | 葡萄糖 Glucose/(g·L-1) | 蚕蛹粉 Soybean cake meal/(g·L-1) | 接种量 Inoculatin amount/% | Y:菌落数 Number of bacterial colonies/(108 cfu·mL-1) |
---|---|---|---|---|
1 | 15 | 20 | 5 | 10.5 |
2 | 25 | 20 | 5 | 49.2 |
3 | 15 | 30 | 5 | 24.9 |
4 | 25 | 30 | 5 | 29.7 |
5 | 15 | 25 | 2.5 | 3.0 |
6 | 25 | 25 | 2.5 | 32.0 |
7 | 15 | 25 | 7.5 | 8.1 |
8 | 25 | 25 | 7.5 | 34.7 |
9 | 20 | 20 | 2.5 | 37.8 |
10 | 20 | 30 | 2.5 | 5.4 |
11 | 20 | 20 | 7.5 | 20.0 |
12 | 20 | 30 | 7.5 | 26.5 |
13 | 20 | 25 | 5 | 44.3 |
14 | 20 | 25 | 5 | 47.6 |
15 | 20 | 25 | 5 | 50.4 |
16 | 20 | 25 | 5 | 59.5 |
17 | 20 | 25 | 5 | 55.5 |
方差来源 Source | 平方和 Sum of squares | 自由度 Degree of freedom | 均方 Mean square | F值 F value | P值 P value | 显著性 Significance |
---|---|---|---|---|---|---|
模型 Model | 4 965.87 | 9 | 551.76 | 17.31 | 0.000 5 | ** |
X1 | 1 227.60 | 1 | 1 227.60 | 38.51 | 0.000 4 | ** |
X4 | 120.13 | 1 | 120.13 | 3.77 | 0.093 4 | |
X10 | 15.40 | 1 | 15.40 | 0.48 | 0.509 4 | |
X1X4 | 287.30 | 1 | 287.30 | 9.01 | 0.019 9 | * |
X1X10 | 1.44 | 1 | 1.44 | 0.045 | 0.837 8 | |
X4X10 | 378.30 | 1 | 378.30 | 11.87 | 0.010 8 | * |
X12 | 703.94 | 1 | 703.94 | 22.08 | 0.002 2 | ** |
X42 | 417.27 | 1 | 417.27 | 13.09 | 0.008 5 | ** |
X102 | 1 532.83 | 1 | 1 532.83 | 48.08 | 0.000 2 | ** |
残差 Residual | 223.16 | 7 | 31.88 | |||
失拟项 Lack of fit | 74.91 | 3 | 24.97 | 0.67 | 0.611 7 | |
纯误差 Pure error | 148.25 | 4 | 37.06 | |||
总和Cor total | 5 189.04 | 16 | ||||
R2 | 0.957 0 | |||||
Radj2 | 0.901 7 |
Table 6 Regression model for response surface analysis of variance
方差来源 Source | 平方和 Sum of squares | 自由度 Degree of freedom | 均方 Mean square | F值 F value | P值 P value | 显著性 Significance |
---|---|---|---|---|---|---|
模型 Model | 4 965.87 | 9 | 551.76 | 17.31 | 0.000 5 | ** |
X1 | 1 227.60 | 1 | 1 227.60 | 38.51 | 0.000 4 | ** |
X4 | 120.13 | 1 | 120.13 | 3.77 | 0.093 4 | |
X10 | 15.40 | 1 | 15.40 | 0.48 | 0.509 4 | |
X1X4 | 287.30 | 1 | 287.30 | 9.01 | 0.019 9 | * |
X1X10 | 1.44 | 1 | 1.44 | 0.045 | 0.837 8 | |
X4X10 | 378.30 | 1 | 378.30 | 11.87 | 0.010 8 | * |
X12 | 703.94 | 1 | 703.94 | 22.08 | 0.002 2 | ** |
X42 | 417.27 | 1 | 417.27 | 13.09 | 0.008 5 | ** |
X102 | 1 532.83 | 1 | 1 532.83 | 48.08 | 0.000 2 | ** |
残差 Residual | 223.16 | 7 | 31.88 | |||
失拟项 Lack of fit | 74.91 | 3 | 24.97 | 0.67 | 0.611 7 | |
纯误差 Pure error | 148.25 | 4 | 37.06 | |||
总和Cor total | 5 189.04 | 16 | ||||
R2 | 0.957 0 | |||||
Radj2 | 0.901 7 |
Fig. 5 Response surface and contour plots of colony number under the interaction of various factorsNote: A, C and E show the response surface of colony number under the interaction of glucose, soybean cake meal and inoculation quantity, respectively; B, D and F show the corresponding contours, respectively.
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