Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (7): 241-253.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1450
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XIE Dong(), WANG Liu-wei, LI Ning-jian, LI Ze-lin, XU Zi-hang, ZHANG Qing-hua()
Received:
2022-11-25
Online:
2023-07-26
Published:
2023-08-17
Contact:
ZHANG Qing-hua
E-mail:2651553709@qq.com;zqh_net@163.com
XIE Dong, WANG Liu-wei, LI Ning-jian, LI Ze-lin, XU Zi-hang, ZHANG Qing-hua. Exploration, Identification and Phosphorus-solubilizing Condition Optimization of a Multifunctional Strain[J]. Biotechnology Bulletin, 2023, 39(7): 241-253.
因素Factors | 水平Level | |
---|---|---|
-1 | +1 | |
A: 葡萄糖浓度Glucose concentration/(g·L-1) | 15.0 | 20.0 |
B: 磷酸三钙浓度Concentration of tricalcium phosphate/(g·L-1) | 5.0 | 7.0 |
C: 硫酸铵浓度Ammonium sulfate concentration/(g·L-1) | 1.0 | 2.0 |
D: 无机盐浓度Concentration of inorganic salts/(g·L-1) | 0.48 | 0.96 |
E: 初始pH Initial pH | 6.0 | 7.0 |
F: 温度Temperature/℃ | 50.0 | 55.0 |
G: 接种量Inoculum quantity/% | 2.0 | 3.0 |
H: 装液量Loaded liquid/mL | 60.0 | 80.0 |
J: 转速Rotating speed /(r·min-1) | 200 | 225 |
Table 1 Factors and levels of Plackett-Burman experiment design
因素Factors | 水平Level | |
---|---|---|
-1 | +1 | |
A: 葡萄糖浓度Glucose concentration/(g·L-1) | 15.0 | 20.0 |
B: 磷酸三钙浓度Concentration of tricalcium phosphate/(g·L-1) | 5.0 | 7.0 |
C: 硫酸铵浓度Ammonium sulfate concentration/(g·L-1) | 1.0 | 2.0 |
D: 无机盐浓度Concentration of inorganic salts/(g·L-1) | 0.48 | 0.96 |
E: 初始pH Initial pH | 6.0 | 7.0 |
F: 温度Temperature/℃ | 50.0 | 55.0 |
G: 接种量Inoculum quantity/% | 2.0 | 3.0 |
H: 装液量Loaded liquid/mL | 60.0 | 80.0 |
J: 转速Rotating speed /(r·min-1) | 200 | 225 |
因素Factor | 水平Level | |||
---|---|---|---|---|
-1 | 0 | +1 | ||
B: 磷酸三钙浓度 Concentration of tricalcium phosphate/(g·L-1) | 5.5 | 6.5 | 7.5 | |
H: 装液量Loaded liquid/mL | 55.0 | 60.0 | 75.0 | |
J: 转速Rotating speed/(r·min-1) | 195 | 207 | 219 |
Table 2 Factors and levels of Box-Behnken experiment design
因素Factor | 水平Level | |||
---|---|---|---|---|
-1 | 0 | +1 | ||
B: 磷酸三钙浓度 Concentration of tricalcium phosphate/(g·L-1) | 5.5 | 6.5 | 7.5 | |
H: 装液量Loaded liquid/mL | 55.0 | 60.0 | 75.0 | |
J: 转速Rotating speed/(r·min-1) | 195 | 207 | 219 |
Fig. 1 Primary screening and secondary screening of high temperature resistant phosphorus-solubilizing strain A: Comparison of phosphorus-solubilizing ability of high-temperature resistant phosphorus-solubilizing bacteria.(Pz, Px1, Px2, Py1, Py2, Py3 and Pb1 respectively refers to the phosphorus solubilization curves of 7 strains of different phosphorus solubilizers obtained from the preliminary experiments, and CK refers to the blank control group with equal amount of sterile water added). B: Phosphorus-solubilizing curve of strain Pb1
指标 Index | 结果 Result |
---|---|
接触酶试验 Catalase test | + |
淀粉水解 Amylohydrolysis | + |
明胶液化 Gelatin liquefaction | - |
尿素酶反应 Urease reaction | + |
甲基红(M.R)试验 Methyl red test | - |
柠檬酸盐利用 Utilization of citrate | - |
吲哚试验 Indole test | + |
葡萄糖发酵 Fermentation of glucose | + |
蔗糖发酵反应 Sucrose fermentation reaction | + |
Table 3 Physiological and biochemical characteristics of strain Pb1
指标 Index | 结果 Result |
---|---|
接触酶试验 Catalase test | + |
淀粉水解 Amylohydrolysis | + |
明胶液化 Gelatin liquefaction | - |
尿素酶反应 Urease reaction | + |
甲基红(M.R)试验 Methyl red test | - |
柠檬酸盐利用 Utilization of citrate | - |
吲哚试验 Indole test | + |
葡萄糖发酵 Fermentation of glucose | + |
蔗糖发酵反应 Sucrose fermentation reaction | + |
Fig. 4 Effects of different medium components on the phosphorus-solubilizaing ability of B. smithii Pb1 Different lower letters indicate significant differences at P < 0.05 level. The same below
实验序号 Experimen No. | A | B | C | D | E | F | G | H | J | 速效磷浓度 Concentration of available P/(mg·L-1) |
---|---|---|---|---|---|---|---|---|---|---|
1 | 15.0 | 7.0 | 2.0 | 0.96 | 50.0 | 6.0 | 2.0 | 80.0 | 200 | 466.46 |
2 | 15.0 | 7.0 | 2.0 | 0.48 | 55.0 | 7.0 | 3.0 | 60.0 | 200 | 532.56 |
3 | 20.0 | 7.0 | 1.0 | 0.48 | 50.0 | 7.0 | 2.0 | 80.0 | 225 | 430.45 |
4 | 15.0 | 7.0 | 1.0 | 0.96 | 55.0 | 6.0 | 3.0 | 80.0 | 225 | 421.71 |
5 | 20.0 | 5.0 | 2.0 | 0.96 | 50.0 | 7.0 | 3.0 | 80.0 | 200 | 388.76 |
6 | 15.0 | 5.0 | 2.0 | 0.48 | 55.0 | 7.0 | 2.0 | 80.0 | 225 | 416.31 |
7 | 20.0 | 7.0 | 2.0 | 0.48 | 50.0 | 6.0 | 3.0 | 60.0 | 225 | 472.63 |
8 | 20.0 | 5.0 | 2.0 | 0.96 | 55.0 | 6.0 | 2.0 | 60.0 | 225 | 397.27 |
9 | 20.0 | 7.0 | 1.0 | 0.96 | 55.0 | 7.0 | 2.0 | 60.0 | 200 | 500.41 |
10 | 15.0 | 5.0 | 1.0 | 0.96 | 50.0 | 7.0 | 3.0 | 60.0 | 225 | 426.59 |
11 | 20.0 | 5.0 | 1.0 | 0.48 | 55.0 | 6.0 | 3.0 | 80.0 | 200 | 417.08 |
12 | 15.0 | 5.0 | 1.0 | 0.48 | 50.0 | 6.0 | 2.0 | 60.0 | 200 | 530.25 |
Table 4 Design and results of Plackett-Burman experiments
实验序号 Experimen No. | A | B | C | D | E | F | G | H | J | 速效磷浓度 Concentration of available P/(mg·L-1) |
---|---|---|---|---|---|---|---|---|---|---|
1 | 15.0 | 7.0 | 2.0 | 0.96 | 50.0 | 6.0 | 2.0 | 80.0 | 200 | 466.46 |
2 | 15.0 | 7.0 | 2.0 | 0.48 | 55.0 | 7.0 | 3.0 | 60.0 | 200 | 532.56 |
3 | 20.0 | 7.0 | 1.0 | 0.48 | 50.0 | 7.0 | 2.0 | 80.0 | 225 | 430.45 |
4 | 15.0 | 7.0 | 1.0 | 0.96 | 55.0 | 6.0 | 3.0 | 80.0 | 225 | 421.71 |
5 | 20.0 | 5.0 | 2.0 | 0.96 | 50.0 | 7.0 | 3.0 | 80.0 | 200 | 388.76 |
6 | 15.0 | 5.0 | 2.0 | 0.48 | 55.0 | 7.0 | 2.0 | 80.0 | 225 | 416.31 |
7 | 20.0 | 7.0 | 2.0 | 0.48 | 50.0 | 6.0 | 3.0 | 60.0 | 225 | 472.63 |
8 | 20.0 | 5.0 | 2.0 | 0.96 | 55.0 | 6.0 | 2.0 | 60.0 | 225 | 397.27 |
9 | 20.0 | 7.0 | 1.0 | 0.96 | 55.0 | 7.0 | 2.0 | 60.0 | 200 | 500.41 |
10 | 15.0 | 5.0 | 1.0 | 0.96 | 50.0 | 7.0 | 3.0 | 60.0 | 225 | 426.59 |
11 | 20.0 | 5.0 | 1.0 | 0.48 | 55.0 | 6.0 | 3.0 | 80.0 | 200 | 417.08 |
12 | 15.0 | 5.0 | 1.0 | 0.48 | 50.0 | 6.0 | 2.0 | 60.0 | 200 | 530.25 |
方差来源 Source | 平方和 Sum of squares | 自由度 Freedom | 均方 Mean square | 效应 Stdized effects | 贡献率 Contribution/% | F值 F value | P值 P value | 等级 Rank |
---|---|---|---|---|---|---|---|---|
Model | 26 763.70 | 9 | 2 973.74 | 20.04 | 0.0484* | |||
A | 2 922.82 | 1 | 2 922.82 | -31.21 | 10.80 | 19.70 | 0.0472* | 5 |
B | 5 123.68 | 1 | 5 123.68 | 41.33 | 18.93 | 34.53 | 0.0278* | 3 |
C | 229.69 | 1 | 229.69 | -8.75 | 0.85 | 1.55 | 0.3395 | 7 |
D | 3 269.64 | 1 | 3 269.64 | -33.01 | 12.08 | 22.03 | 0.0425* | 4 |
E | 74.00 | 1 | 74.00 | -4.97 | 0.27 | 0.50 | 0.5333 | 8 |
F | 8.88 | 1 | 8.88 | -1.72 | 0.03 | 0.06 | 0.8296 | 9 |
G | 557.88 | 1 | 557.88 | -13.64 | 2.06 | 3.76 | 0.1921 | 6 |
H | 8 476.89 | 1 | 8 476.89 | -53.16 | 31.33 | 57.12 | 0.0171* | 1 |
J | 6 100.23 | 1 | 6 100.23 | -45.09 | 22.54 | 41.11 | 0.0235* | 2 |
Table 5 Variance analysis of Plackett-Burman experiments results
方差来源 Source | 平方和 Sum of squares | 自由度 Freedom | 均方 Mean square | 效应 Stdized effects | 贡献率 Contribution/% | F值 F value | P值 P value | 等级 Rank |
---|---|---|---|---|---|---|---|---|
Model | 26 763.70 | 9 | 2 973.74 | 20.04 | 0.0484* | |||
A | 2 922.82 | 1 | 2 922.82 | -31.21 | 10.80 | 19.70 | 0.0472* | 5 |
B | 5 123.68 | 1 | 5 123.68 | 41.33 | 18.93 | 34.53 | 0.0278* | 3 |
C | 229.69 | 1 | 229.69 | -8.75 | 0.85 | 1.55 | 0.3395 | 7 |
D | 3 269.64 | 1 | 3 269.64 | -33.01 | 12.08 | 22.03 | 0.0425* | 4 |
E | 74.00 | 1 | 74.00 | -4.97 | 0.27 | 0.50 | 0.5333 | 8 |
F | 8.88 | 1 | 8.88 | -1.72 | 0.03 | 0.06 | 0.8296 | 9 |
G | 557.88 | 1 | 557.88 | -13.64 | 2.06 | 3.76 | 0.1921 | 6 |
H | 8 476.89 | 1 | 8 476.89 | -53.16 | 31.33 | 57.12 | 0.0171* | 1 |
J | 6 100.23 | 1 | 6 100.23 | -45.09 | 22.54 | 41.11 | 0.0235* | 2 |
试验序号 Experiment No. | B | H | J | 速效磷浓度 Concentration of available P/(mg·L-1) |
---|---|---|---|---|
1 | 5.0 | 80.0 | 225 | 459.26 |
2 | 5.5 | 75.0 | 219 | 487.29 |
3 | 6.0 | 70.0 | 213 | 516.10 |
4 | 6.5 | 65.0 | 207 | 533.28 |
5 | 7.0 | 60.0 | 200 | 529.48 |
6 | 7.5 | 55.0 | 193 | 508.67 |
Table 6 Design and results of the steepest ascent test
试验序号 Experiment No. | B | H | J | 速效磷浓度 Concentration of available P/(mg·L-1) |
---|---|---|---|---|
1 | 5.0 | 80.0 | 225 | 459.26 |
2 | 5.5 | 75.0 | 219 | 487.29 |
3 | 6.0 | 70.0 | 213 | 516.10 |
4 | 6.5 | 65.0 | 207 | 533.28 |
5 | 7.0 | 60.0 | 200 | 529.48 |
6 | 7.5 | 55.0 | 193 | 508.67 |
方差来源Source | 平方和Sum of squares | 自由度Freedom | 均方Mean square | F值F value | P值P value | 显著性Significance |
---|---|---|---|---|---|---|
Model | 12 744.38 | 9 | 1 416.04 | 29.54 | 0.000 8 | ** |
B | 513.60 | 1 | 513.60 | 10.72 | 0.022 1 | * |
H | 39.60 | 1 | 39.60 | 0.83 | 0.405 0 | |
J | 6.48 | 1 | 6.48 | 0.14 | 0.728 2 | |
BH | 88.17 | 1 | 88.17 | 1.84 | 0.233 0 | |
BJ | 122.99 | 1 | 122.99 | 2.57 | 0.170 1 | |
HJ | 258.24 | 1 | 258.24 | 5.39 | 0.068 0 | |
B2 | 5 892.93 | 1 | 5 892.93 | 122.94 | 0.000 1 | ** |
H2 | 4 762.66 | 1 | 4 762.66 | 99.36 | 0.000 2 | ** |
J2 | 2 787.23 | 1 | 2 787.23 | 58.15 | 0.000 6 | ** |
残差 Residual | 239.66 | 5 | 49.93 | |||
失拟性 Lack of fit | 225.63 | 3 | 75.21 | 10.72 | 0.086 5 | |
净误差 Pure error | 14.03 | 2 | 7.01 | |||
合计 Total | 12 984.04 | 14 | ||||
决定系数 R2 | 0.981 5 | |||||
校正系数R2 adj | 0.948 3 | |||||
Pred R2 | 0.719 5 | |||||
Adeq precision | 16.062 |
Table 7 Variance analysis of regression model
方差来源Source | 平方和Sum of squares | 自由度Freedom | 均方Mean square | F值F value | P值P value | 显著性Significance |
---|---|---|---|---|---|---|
Model | 12 744.38 | 9 | 1 416.04 | 29.54 | 0.000 8 | ** |
B | 513.60 | 1 | 513.60 | 10.72 | 0.022 1 | * |
H | 39.60 | 1 | 39.60 | 0.83 | 0.405 0 | |
J | 6.48 | 1 | 6.48 | 0.14 | 0.728 2 | |
BH | 88.17 | 1 | 88.17 | 1.84 | 0.233 0 | |
BJ | 122.99 | 1 | 122.99 | 2.57 | 0.170 1 | |
HJ | 258.24 | 1 | 258.24 | 5.39 | 0.068 0 | |
B2 | 5 892.93 | 1 | 5 892.93 | 122.94 | 0.000 1 | ** |
H2 | 4 762.66 | 1 | 4 762.66 | 99.36 | 0.000 2 | ** |
J2 | 2 787.23 | 1 | 2 787.23 | 58.15 | 0.000 6 | ** |
残差 Residual | 239.66 | 5 | 49.93 | |||
失拟性 Lack of fit | 225.63 | 3 | 75.21 | 10.72 | 0.086 5 | |
净误差 Pure error | 14.03 | 2 | 7.01 | |||
合计 Total | 12 984.04 | 14 | ||||
决定系数 R2 | 0.981 5 | |||||
校正系数R2 adj | 0.948 3 | |||||
Pred R2 | 0.719 5 | |||||
Adeq precision | 16.062 |
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