Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (8): 299-308.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0040
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ZHANG Meng-fei1(), YU Lian1(), LI Fei2(), LI Zhe2, SU Xin-ying2, LAN Cai-bi2, ZHU Hu1, QIN Ying1
Received:
2024-01-12
Online:
2024-08-26
Published:
2024-07-31
Contact:
YU Lian, LI Fei
E-mail:1913525246@qq.com;lianyu@gxu.edu.cn;finylee@yeah.net
ZHANG Meng-fei, YU Lian, LI Fei, LI Zhe, SU Xin-ying, LAN Cai-bi, ZHU Hu, QIN Ying. Optimization of Fermentation Culture Conditions for Macrolactins Yield of Bacillus siamensis Using Response Surface Methodology[J]. Biotechnology Bulletin, 2024, 40(8): 299-308.
因素Factor/(g·L-1) | 低水平Low level(-1) | 高水平High level(1) |
---|---|---|
葡萄糖(X1) | 10 | 20 |
酵母提取物(X2) | 0.6 | 1.0 |
(NH4)2SO4(X3) | 1.0 | 3.0 |
缬氨酸(X4) | 2.0 | 6.0 |
组氨酸(X5) | 2.0 | 6.0 |
Table 1 Factors and levels of experiment designed by Plackett-Burman(PBD)
因素Factor/(g·L-1) | 低水平Low level(-1) | 高水平High level(1) |
---|---|---|
葡萄糖(X1) | 10 | 20 |
酵母提取物(X2) | 0.6 | 1.0 |
(NH4)2SO4(X3) | 1.0 | 3.0 |
缬氨酸(X4) | 2.0 | 6.0 |
组氨酸(X5) | 2.0 | 6.0 |
水平 Level | 葡萄糖Glucose X1/(g·L-1) | 酵母提取物Yeast extract X2/(g·L-1) | 缬氨酸Valine X4/(g·L-1) | 组氨酸Histidine X5/(g·L-1) |
---|---|---|---|---|
-1 | 15 | 0.8 | 5.0 | 5.0 |
0 | 20 | 1.0 | 6.0 | 6.0 |
1 | 25 | 1.2 | 7.0 | 7.0 |
Table 2 Factors and levels of response surface tests design
水平 Level | 葡萄糖Glucose X1/(g·L-1) | 酵母提取物Yeast extract X2/(g·L-1) | 缬氨酸Valine X4/(g·L-1) | 组氨酸Histidine X5/(g·L-1) |
---|---|---|---|---|
-1 | 15 | 0.8 | 5.0 | 5.0 |
0 | 20 | 1.0 | 6.0 | 6.0 |
1 | 25 | 1.2 | 7.0 | 7.0 |
序号Number | 葡萄糖Glucose X1 | 酵母提取物Yeast extract X2 | (NH4)2SO4 X3 | 缬氨酸Valine X4 | 组氨酸Histidine X5 | MLNs产量MLNs yield×102/(μg·mL-1) |
---|---|---|---|---|---|---|
1 | 1 | -1 | 1 | 1 | -1 | 3.475 |
2 | 1 | 1 | -1 | 1 | -1 | 3.292 |
3 | -1 | 1 | 1 | 1 | 1 | 3.196 |
4 | 1 | -1 | -1 | -1 | 1 | 3.508 |
5 | 1 | 1 | -1 | 1 | 1 | 3.213 |
6 | -1 | -1 | -1 | 1 | -1 | 3.454 |
7 | 1 | -1 | 1 | -1 | 1 | 3.588 |
8 | -1 | -1 | 1 | 1 | 1 | 3.279 |
9 | -1 | 1 | 1 | -1 | -1 | 3.400 |
10 | -1 | 1 | -1 | -1 | 1 | 3.254 |
11 | 1 | 1 | 1 | -1 | -1 | 3.413 |
12 | -1 | -1 | -1 | -1 | -1 | 3.500 |
Table 3 Design and results of PBD
序号Number | 葡萄糖Glucose X1 | 酵母提取物Yeast extract X2 | (NH4)2SO4 X3 | 缬氨酸Valine X4 | 组氨酸Histidine X5 | MLNs产量MLNs yield×102/(μg·mL-1) |
---|---|---|---|---|---|---|
1 | 1 | -1 | 1 | 1 | -1 | 3.475 |
2 | 1 | 1 | -1 | 1 | -1 | 3.292 |
3 | -1 | 1 | 1 | 1 | 1 | 3.196 |
4 | 1 | -1 | -1 | -1 | 1 | 3.508 |
5 | 1 | 1 | -1 | 1 | 1 | 3.213 |
6 | -1 | -1 | -1 | 1 | -1 | 3.454 |
7 | 1 | -1 | 1 | -1 | 1 | 3.588 |
8 | -1 | -1 | 1 | 1 | 1 | 3.279 |
9 | -1 | 1 | 1 | -1 | -1 | 3.400 |
10 | -1 | 1 | -1 | -1 | 1 | 3.254 |
11 | 1 | 1 | 1 | -1 | -1 | 3.413 |
12 | -1 | -1 | -1 | -1 | -1 | 3.500 |
来源Source | 自由度Freedom | 离均差平方和Sum of squares of deviation from mean | 均方差Standard deviation | F值 F value | P值P value |
---|---|---|---|---|---|
模型 | 5 | 0.170 | 0.034 | 15.76 | 0.002 1 |
X1 | 1 | 0.014 | 0.014 | 6.28 | 0.046 2 |
X2 | 1 | 0.089 | 0.089 | 40.88 | 0.000 7 |
X3 | 1 | 0.001 | 0.001 | 0.64 | 0.453 0 |
X4 | 1 | 0.047 | 0.047 | 21.65 | 0.003 5 |
X5 | 1 | 0.021 | 0.021 | 9.37 | 0.022 2 |
残差 | 6 | 0.013 | 0.002 | ||
总离差 | 11 | 0.190 |
Table 4 Variance analysis of PBD results
来源Source | 自由度Freedom | 离均差平方和Sum of squares of deviation from mean | 均方差Standard deviation | F值 F value | P值P value |
---|---|---|---|---|---|
模型 | 5 | 0.170 | 0.034 | 15.76 | 0.002 1 |
X1 | 1 | 0.014 | 0.014 | 6.28 | 0.046 2 |
X2 | 1 | 0.089 | 0.089 | 40.88 | 0.000 7 |
X3 | 1 | 0.001 | 0.001 | 0.64 | 0.453 0 |
X4 | 1 | 0.047 | 0.047 | 21.65 | 0.003 5 |
X5 | 1 | 0.021 | 0.021 | 9.37 | 0.022 2 |
残差 | 6 | 0.013 | 0.002 | ||
总离差 | 11 | 0.190 |
试验编号No. | 葡萄糖Glucose X1/(g·L-1) | 酵母提取物Yeast extract X2/(g·L-1) | 缬氨酸Valine X4/(g·L-1) | 组氨酸Histidine X5/(g·L-1) | MLNs产量MLNs yield×102/(μg·mL-1) |
---|---|---|---|---|---|
1 | 25.0 | 1.0 | 5.0 | 6.0 | 3.13 |
2 | 15.0 | 1.0 | 6.0 | 5.0 | 2.85 |
3 | 20.0 | 0.8 | 6.0 | 7.0 | 2.96 |
4 | 25.0 | 0.8 | 6.0 | 6.0 | 4.01 |
5 | 20.0 | 1.0 | 6.0 | 6.0 | 5.01 |
6 | 20.0 | 1.0 | 6.0 | 6.0 | 5.43 |
7 | 20.0 | 1.2 | 6.0 | 5.0 | 3.57 |
8 | 20.0 | 1.2 | 5.0 | 6.0 | 3.08 |
9 | 20.0 | 0.8 | 7.0 | 6.0 | 3.32 |
10 | 20.0 | 1.0 | 7.0 | 5.0 | 4.07 |
11 | 20.0 | 1.0 | 6.0 | 6.0 | 5.37 |
12 | 15.0 | 1.2 | 6.0 | 6.0 | 2.98 |
13 | 20.0 | 0.8 | 5.0 | 6.0 | 2.96 |
14 | 15.0 | 1.0 | 5.0 | 6.0 | 2.90 |
15 | 20.0 | 1.2 | 7.0 | 6.0 | 3.79 |
16 | 15.0 | 0.8 | 6.0 | 6.0 | 3.42 |
17 | 20.0 | 1.0 | 5.0 | 7.0 | 3.41 |
18 | 15.0 | 1.0 | 6.0 | 7.0 | 3.15 |
19 | 20.0 | 0.8 | 6.0 | 5.0 | 3.09 |
20 | 20.0 | 1.0 | 5.0 | 5.0 | 3.39 |
21 | 20.0 | 1.2 | 6.0 | 7.0 | 2.99 |
22 | 20.0 | 1.0 | 7.0 | 7.0 | 3.72 |
23 | 15.0 | 1.0 | 7.0 | 6.0 | 2.58 |
24 | 20.0 | 1.0 | 6.0 | 6.0 | 5.30 |
25 | 25.0 | 1.2 | 6.0 | 6.0 | 4.54 |
26 | 20.0 | 1.0 | 6.0 | 6.0 | 5.41 |
27 | 25.0 | 1.0 | 7.0 | 6.0 | 4.45 |
28 | 25.0 | 1.0 | 6.0 | 7.0 | 4.07 |
29 | 25.0 | 1.0 | 6.0 | 5.0 | 4.28 |
Table 5 Response surface test for optimal yield of MLNs from B. siamensis strain K53
试验编号No. | 葡萄糖Glucose X1/(g·L-1) | 酵母提取物Yeast extract X2/(g·L-1) | 缬氨酸Valine X4/(g·L-1) | 组氨酸Histidine X5/(g·L-1) | MLNs产量MLNs yield×102/(μg·mL-1) |
---|---|---|---|---|---|
1 | 25.0 | 1.0 | 5.0 | 6.0 | 3.13 |
2 | 15.0 | 1.0 | 6.0 | 5.0 | 2.85 |
3 | 20.0 | 0.8 | 6.0 | 7.0 | 2.96 |
4 | 25.0 | 0.8 | 6.0 | 6.0 | 4.01 |
5 | 20.0 | 1.0 | 6.0 | 6.0 | 5.01 |
6 | 20.0 | 1.0 | 6.0 | 6.0 | 5.43 |
7 | 20.0 | 1.2 | 6.0 | 5.0 | 3.57 |
8 | 20.0 | 1.2 | 5.0 | 6.0 | 3.08 |
9 | 20.0 | 0.8 | 7.0 | 6.0 | 3.32 |
10 | 20.0 | 1.0 | 7.0 | 5.0 | 4.07 |
11 | 20.0 | 1.0 | 6.0 | 6.0 | 5.37 |
12 | 15.0 | 1.2 | 6.0 | 6.0 | 2.98 |
13 | 20.0 | 0.8 | 5.0 | 6.0 | 2.96 |
14 | 15.0 | 1.0 | 5.0 | 6.0 | 2.90 |
15 | 20.0 | 1.2 | 7.0 | 6.0 | 3.79 |
16 | 15.0 | 0.8 | 6.0 | 6.0 | 3.42 |
17 | 20.0 | 1.0 | 5.0 | 7.0 | 3.41 |
18 | 15.0 | 1.0 | 6.0 | 7.0 | 3.15 |
19 | 20.0 | 0.8 | 6.0 | 5.0 | 3.09 |
20 | 20.0 | 1.0 | 5.0 | 5.0 | 3.39 |
21 | 20.0 | 1.2 | 6.0 | 7.0 | 2.99 |
22 | 20.0 | 1.0 | 7.0 | 7.0 | 3.72 |
23 | 15.0 | 1.0 | 7.0 | 6.0 | 2.58 |
24 | 20.0 | 1.0 | 6.0 | 6.0 | 5.30 |
25 | 25.0 | 1.2 | 6.0 | 6.0 | 4.54 |
26 | 20.0 | 1.0 | 6.0 | 6.0 | 5.41 |
27 | 25.0 | 1.0 | 7.0 | 6.0 | 4.45 |
28 | 25.0 | 1.0 | 6.0 | 7.0 | 4.07 |
29 | 25.0 | 1.0 | 6.0 | 5.0 | 4.28 |
来源Source | 平方和Sum of squares | 自由度Freedom | 均方Meansquare | F值F value | P值P value |
---|---|---|---|---|---|
模型 | 20.19 | 14 | 1.44 | 17.10 | < 0.000 1 |
A-葡萄糖 | 3.63 | 1 | 3.63 | 43.04 | < 0.000 1 |
B-酵母提取物 | 0.12 | 1 | 0.12 | 1.40 | 0.256 6 |
C-缬氨酸 | 0.78 | 1 | 0.78 | 9.25 | 0.008 8 |
D-组氨酸 | 0.08 | 1 | 0.08 | 0.89 | 0.361 0 |
AB | 0.24 | 1 | 0.24 | 2.79 | 0.117 1 |
AC | 0.67 | 1 | 0.67 | 7.97 | 0.013 5 |
AD | 0.06 | 1 | 0.06 | 0.77 | 0.394 7 |
BC | 0.03 | 1 | 0.03 | 0.36 | 0.556 4 |
BD | 0.05 | 1 | 0.05 | 0.60 | 0.451 4 |
CD | 0.03 | 1 | 0.03 | 0.41 | 0.534 4 |
A2 | 4.19 | 1 | 4.19 | 49.62 | < 0.000 1 |
B2 | 6.61 | 1 | 6.61 | 78.38 | < 0.000 1 |
C2 | 6.46 | 1 | 6.46 | 76.64 | < 0.000 1 |
D2 | 5.31 | 1 | 5.31 | 62.94 | < 0.000 1 |
误差 | 1.18 | 14 | 0.08 | ||
失拟项 | 1.06 | 10 | 0.11 | 3.61 | 0.114 0 |
纯误差 | 0.12 | 4 | 0.0.3 | ||
总和 | 21.37 | 28 |
Table 6 Variance analysis of regression model
来源Source | 平方和Sum of squares | 自由度Freedom | 均方Meansquare | F值F value | P值P value |
---|---|---|---|---|---|
模型 | 20.19 | 14 | 1.44 | 17.10 | < 0.000 1 |
A-葡萄糖 | 3.63 | 1 | 3.63 | 43.04 | < 0.000 1 |
B-酵母提取物 | 0.12 | 1 | 0.12 | 1.40 | 0.256 6 |
C-缬氨酸 | 0.78 | 1 | 0.78 | 9.25 | 0.008 8 |
D-组氨酸 | 0.08 | 1 | 0.08 | 0.89 | 0.361 0 |
AB | 0.24 | 1 | 0.24 | 2.79 | 0.117 1 |
AC | 0.67 | 1 | 0.67 | 7.97 | 0.013 5 |
AD | 0.06 | 1 | 0.06 | 0.77 | 0.394 7 |
BC | 0.03 | 1 | 0.03 | 0.36 | 0.556 4 |
BD | 0.05 | 1 | 0.05 | 0.60 | 0.451 4 |
CD | 0.03 | 1 | 0.03 | 0.41 | 0.534 4 |
A2 | 4.19 | 1 | 4.19 | 49.62 | < 0.000 1 |
B2 | 6.61 | 1 | 6.61 | 78.38 | < 0.000 1 |
C2 | 6.46 | 1 | 6.46 | 76.64 | < 0.000 1 |
D2 | 5.31 | 1 | 5.31 | 62.94 | < 0.000 1 |
误差 | 1.18 | 14 | 0.08 | ||
失拟项 | 1.06 | 10 | 0.11 | 3.61 | 0.114 0 |
纯误差 | 0.12 | 4 | 0.0.3 | ||
总和 | 21.37 | 28 |
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