Optimization for the Production of Deoxynivalenoland Zearalenone by Fusarium graminearum UsingResponse Surface Methodology
Abstract
:1. Introduction
2. Results and Discussion
2.1. Model Fitting and Statistical Analysis
2.2. Response Surface Analysis
2.3. Optimization of Independent Variables and Validation of the Model
2.4. Purification and Purity of Mycotoxin Samples for Potential Analytical Standards
3. Conclusions
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Fungal and Inoculum Preparation
4.3. Culture Conditions for Mycotoxin Production of F.graminearum Strain 29
4.4. Experimental Design for Mycotoxin Production of F. graminearum Strain 29
4.5. Mycotoxin Determination
4.6. Mycotoxins Extraction, Purification and Analysis for Potential Analyte Standards
4.6.1. Mycotoxin Extraction from Growth Medium
4.6.2. Mycotoxin Seperation and Purification by Preparative Column HPLC
4.6.3. Identification and Purity of the Isolated Mycotoxin Samples
4.7. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
References
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No. | X1 | X2 | X3 | Mycotoxin Levles (ng/mL) | |
---|---|---|---|---|---|
PH | Temperature (°C) | Time (day) | DON | ZEA | |
1 | −1 (3) | −1 (15) | 0 (21) | 961.02 ± 35.78 | 419.97 ± 30.25 |
2 | 1 (9) | −1 (15) | 0 (21) | <LOD | 19084.20 ± 1102.56 |
3 | −1 (3) | 1 (25) | 0 (21) | 1493.05 ± 91.23 | 609.66 ± 41.35 |
4 | 1 (9) | 1 (25) | 0 (21) | <LOD | 13457.90 ± 978.25 |
5 | −1 (3) | 0 (20) | −1 (14) | 585.59 ± 39.75 | 818.34 ± 54.25 |
6 | 1 (9) | 0 (20) | −1 (14) | <LOD | 13941.4 ± 886.14 |
7 | −1 (3) | 0 (20) | 1 (28) | 2003.54 ± 123.47 | 2288.42 ± 231.52 |
8 | 1 (9) | 0 (20) | 1 (28) | <LOD | 20047.30 ± 1203.59 |
9 | 0 (6) | −1 (15) | −1 (14) | 1559.09 ± 101.27 | 4118.31 ± 334.59 |
10 | 0 (6) | 1 (25) | −1 (14) | 1405.5 ± 89.58 | 2259.09 ± 189.67 |
11 | 0 (6) | −1 (15) | 1 (28) | 2098.03 ± 128.71 | 8661.47 ± 678.15 |
12 | 0 (6) | 1 (25) | 1 (28) | 2605.26 ± 134.55 | 5319.75 ± 408.14 |
13 | 0 (6) | 0 (20) | 0 (21) | 2276.33 ± 121.30 | 3198.34 ± 278.27 |
14 | 0 (6) | 0 (20) | 0 (21) | 2313.33 ± 108.24 | 3133.31 ± 256.77 |
15 | 0 (6) | 0 (20) | 0 (21) | 2332.08 ± 107.57 | 3126.45 ± 281.73 |
16 | 0 (6) | 0 (20) | 0 (21) | 2283.66 ± 119.35 | 3156.87 ± 312.25 |
17 | 0 (6) | 0 (20) | 0 (21) | 2376.34 ± 123.83 | 3185.81 ± 243.51 |
Source | Sum of Squares | df | Mean Squares | F-Value | p-Value |
---|---|---|---|---|---|
DON | |||||
Model | 1.520 × 107 | 9 | 1.689 × 106 | 461.94 | 0.0001 |
X1 | 3.179 × 106 | 1 | 3.179 × 106 | 869.41 | <0.0001 |
X2 | 98051.42 | 1 | 98051.42 | 26.82 | 0.0013 |
X3 | 1.246 × 106 | 1 | 1.246 × 106 | 340.77 | 0.0001 |
X1 X2 | 70763.98 | 1 | 70763.98 | 19.35 | 0.0032 |
X1 X3 | 5.030 × 105 | 1 | 5.030 × 105 | 137.57 | <0.0001 |
X2 X3 | 1.092 × 105 | 1 | 1.092 × 105 | 29.86 | 0.0009 |
X12 | 9.302 × 106 | 1 | 9.302 × 106 | 2544.13 | <0.0001 |
X22 | 1.974 × 105 | 1 | 1.974 × 105 | 53.99 | 0.0002 |
X32 | 1.408 × 105 | 1 | 1.408 × 105 | 38.51 | 0.0005 |
Residual | 25592.79 | 7 | 3656.11 | ||
Lack of Fit | 19068.00 | 3 | 6356.00 | 3.90 | 0.1110 |
Pure Error | 6524.79 | 4 | 1631.20 | ||
Cor Total | 1.523 × 107 | 16 | |||
ZEN | |||||
Model | 6.534 × 108 | 9 | 7.260 × 107 | 2884.71 | <0.0001 |
X1 | 4.882 × 108 | 1 | 4.882 × 108 | 19398.73 | 0.0001 |
X2 | 1.414 × 107 | 1 | 1.414 × 107 | 562.05 | 0.0001 |
X3 | 2.918 × 107 | 1 | 2.918 × 107 | 1159.65 | 0.0001 |
X1 X2 | 7.885 × 106 | 1 | 7.885 × 106 | 313.31 | 0.0001 |
X1 X3 | 5.607 × 106 | 1 | 5.607 × 106 | 222.80 | 0.0001 |
X2 X3 | 5.495 × 105 | 1 | 5.495 × 105 | 21.83 | 0.0023 |
X12 | 9.285 × 107 | 1 | 9.285 × 107 | 3689.54 | 0.0001 |
X22 | 9.977 × 105 | 1 | 9.977 × 105 | 39.65 | 0.0004 |
X32 | 8.764 × 106 | 1 | 8.764 × 106 | 348.24 | 0.0001 |
Residual | 1.762 × 105 | 7 | 25166.28 | ||
Lack of Fit | 175500 | 3 | 57393.40 | 5.76 | 0.0710 |
Pure Error | 3983.75 | 4 | 995.94 | ||
Cor Total | 6.536 × 108 | 16 |
Variables | Symbols | Range and Levels | ||
---|---|---|---|---|
Low (−1) | Medium (0) | High (+1) | ||
Temperature (°C) | X1 | 15 | 20 | 25 |
pH | X2 | 3 | 6 | 9 |
Time (d) | X3 | 14 | 21 | 28 |
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Wu, L.; Qiu, L.; Zhang, H.; Sun, J.; Hu, X.; Wang, B. Optimization for the Production of Deoxynivalenoland Zearalenone by Fusarium graminearum UsingResponse Surface Methodology. Toxins 2017, 9, 57. https://doi.org/10.3390/toxins9020057
Wu L, Qiu L, Zhang H, Sun J, Hu X, Wang B. Optimization for the Production of Deoxynivalenoland Zearalenone by Fusarium graminearum UsingResponse Surface Methodology. Toxins. 2017; 9(2):57. https://doi.org/10.3390/toxins9020057
Chicago/Turabian StyleWu, Li, Lijuan Qiu, Huijie Zhang, Juan Sun, Xuexu Hu, and Bujun Wang. 2017. "Optimization for the Production of Deoxynivalenoland Zearalenone by Fusarium graminearum UsingResponse Surface Methodology" Toxins 9, no. 2: 57. https://doi.org/10.3390/toxins9020057