Production and Characterization of Fengycin by Indigenous Bacillus subtilis F29-3 Originating from a Potato Farm
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of Fengycin
2.1.1. IR Spectrometric Analyses
2.1.2. UV Spectrometric Analyses
2.1.3. NMR Spectrometric Analyses
2.1.4. MALDI-TOF/MASS Analyses
2.2. Optimization of Medium Constituents for Fengycin Production by RSM
2.2.1. Fractional Factorial Design
2.2.2. Steepest Ascent Method
2.2.3. Response Surface Methodology (RSM)
3. Experimental Section
3.1. Microorganism
3.2. Growth Medium and Culture Conditions
3.3. Purification of Fengycin
3.4. Characterization of Fengycin
3.4.1. Fast Atom Bombardment-Mass Spectrometry (FAB-MS)
3.4.2. IR Spectrometric Analyses
3.4.3. UV Spectrometric Analyses
3.4.4. NMR Analysis
3.4.5. MALDI-TOF/MASS
3.5. Quantification of Fengycin
3.6. Optimization of Medium Composition for Fengycin Production
3.6.1. Fractional Factorial Design
3.6.2. Steepest Ascent Method
3.6.3. Response Surface Methodology (RSM)
4. Conclusions
Supplementary Material
ijms-11-04526-s001.pdfAcknowledgments
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Retention Time (min) | Main MALDI-TOF Peak(s) (m/z) | Assignment |
---|---|---|
5, 6 | 1523.865, 1509.855 | B-C16 and C17 fengycin [M + Na]+ |
6, 7 | 1509.855, 1477.828, 1491.825 | B-C16 fengycin [M + Na]+ |
A-C17 fengycin [M + H]+ | ||
B-C16 fengycin [M + H]+ | ||
7, 8 | 1491.825, 1505.851 | B-C16 and C17 fengycin [M + H]+ |
8, 9 | 1505.898, 1527.901 | B-C17 fengycin [M + H]+ |
B-C17 fengycin [M + Na]+ | ||
9, 10 | 1475.844 | A-C17 fengycin [M + H]+ |
10, 11 | 1475.852, 1497.859 | A-C17 fengycin [M + H]+ |
A-C17 fengycin [M + Na]+ | ||
11, 12 | 1475.817, 1497.816 | A-C17 fengycin [M + H]+ |
A-C17 fengycin [M + Na]+ | ||
12, 13 | 1475.793, 1505.808 | A-C17 fengycin [M + H]+ |
B-C17 fengycin [M + H]+ | ||
13, 14 | 1511.853 | B-C16 fengycin [M + Na]+ |
14, 15 | 1489.836 | B-C16 fengycin [M + H]+ |
15, 16 | 1489.912 | B-C16 fengycin [M + H]+ |
Run No. | Variables | |||||||
---|---|---|---|---|---|---|---|---|
Mannitol (g/100 mL) | Soybean Meal (g/100 mL) | NaNO3 (g/100 mL) | FeCl2·4H2O (g/100 mL) | MgSO4·7H2O (g/100 mL) | MnSO4·4H2O (g/100 mL) | Na2MoO4 (g/100 mL) | Fengycin Production (mg/L) | |
1 | −1 | −1 | −1 | −1 | −1 | −1 | −1 | 337 ± 31 |
2 | −1 | −1 | −1 | 1 | 1 | 1 | 1 | 1161± 104 |
3 | −1 | −1 | 1 | −1 | 1 | 1 | −1 | 708 ± 63 |
4 | −1 | −1 | 1 | 1 | −1 | −1 | 1 | 542 ± 72 |
5 | −1 | 1 | −1 | −1 | 1 | −1 | 1 | 447 ± 51 |
6 | −1 | 1 | −1 | 1 | −1 | 1 | −1 | 1688 ± 137 |
7 | −1 | 1 | 1 | −1 | −1 | 1 | 1 | 1066 ± 101 |
8 | −1 | 1 | 1 | 1 | 1 | −1 | −1 | 644 ± 75 |
9 | 1 | −1 | −1 | −1 | −1 | 1 | 1 | 1712 ± 148 |
10 | 1 | −1 | −1 | 1 | 1 | −1 | −1 | 1598 ± 193 |
11 | 1 | −1 | 1 | −1 | 1 | −1 | 1 | 1054 ± 119 |
12 | 1 | −1 | 1 | 1 | −1 | 1 | −1 | 1527 ± 124 |
13 | 1 | 1 | −1 | −1 | 1 | 1 | −1 | 2311 ±254 |
14 | 1 | 1 | −1 | 1 | −1 | −1 | 1 | 2527 ± 285 |
15 | 1 | 1 | 1 | −1 | −1 | −1 | −1 | 1556 ± 199 |
16 | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1853 ± 162 |
Source | DF | Sum of Squares | F-Ratio | Prob > F |
---|---|---|---|---|
Model | 7 | 5925107.9 | 11.7 | 0.0012 |
Mannitol | 1 | 3557939.1 | 49.1 | 0.0001 |
Soybean meal | 1 | 745200.6 | 10.3 | 0.0125 |
NaNO3 | 1 | 500910.1 | 6.9 | 0.0302 |
FeCl2·4H2O | 1 | 344862.6 | 4.8 | 0.0606 |
MgSO4·7H2O | 1 | 86877.6 | 1.2 | 0.3052 |
MnSO4·4H2O | 1 | 689315.1 | 9.5 | 0.0150 |
Na2MoO4 | 1 | 3.1 | 0.0 | 0.9950 |
Experiment No. | Mannitol (g/100 mL) | Soybean Meal (g/100 mL) | NaNO3 (g/100 mL) | MnSO4·4H2O (g/100 mL) |
---|---|---|---|---|
4 | 3.2 | 2.3 | 0.2 | 0.01 |
3 | 2.7 | 2.1 | 0.3 | 0.02 |
2 | 2.3 | 1.9 | 0.4 | 0.03 |
1 | 1.8 | 1.6 | 0.5 | 0.04 |
0 | 1.4 | 1.4 | 0.6 | 0.05 |
−1 | 1.0 | 1.2 | 0.7 | 0.06 |
−2 | 0.5 | 0.9 | 0.8 | 0.07 |
Run No. | Mannitol (g/100 mL) | Soybean Meal (g/100 mL) | NaNO3 (g/100 mL) | MnSO4·4H2O (g/100 mL) | Fengycin Production (mg/L) | |
---|---|---|---|---|---|---|
Experimental | Predicted | |||||
1 | −1 | −1 | −1 | −1 | 3033 ± 313 | 2956 ± 315 |
2 | −1 | −1 | −1 | 1 | 2394 ± 259 | 2517 ± 281 |
3 | −1 | −1 | 1 | −1 | 2461 ± 216 | 2218 ± 215 |
4 | −1 | −1 | 1 | 1 | 1981 ± 238 | 1941 ± 221 |
5 | −1 | 1 | −1 | −1 | 3351 ± 315 | 3327 ± 381 |
6 | −1 | 1 | −1 | 1 | 2699 ± 289 | 2807 ± 252 |
7 | −1 | 1 | 1 | −1 | 2682 ± 278 | 2909 ± 264 |
8 | −1 | 1 | 1 | 1 | 2623 ± 242 | 2550 ± 281 |
9 | 1 | −1 | −1 | −1 | 2867 ± 236 | 2938 ± 312 |
10 | 1 | −1 | −1 | 1 | 2968 ± 256 | 2695 ± 261 |
11 | 1 | −1 | 1 | −1 | 2613 ± 281 | 2516 ± 274 |
12 | 1 | −1 | 1 | 1 | 2414 ± 261 | 2435 ± 253 |
13 | 1 | 1 | −1 | −1 | 2858 ± 275 | 2909 ± 287 |
14 | 1 | 1 | −1 | 1 | 2343 ± 284 | 2584 ± 261 |
15 | 1 | 1 | 1 | −1 | 2933 ± 213 | 2807 ± 271 |
16 | 1 | 1 | 1 | 1 | 2554 ± 215 | 2643 ± 284 |
17 | 0 | 0 | 0 | 0 | 3263 ± 326 | 3371 ± 391 |
18 | 0 | 0 | 0 | 0 | 3418 ± 321 | 3371 ± 337 |
19 | 0 | 0 | 0 | 0 | 3297 ± 323 | 3371 ± 312 |
20 | 0 | 0 | 0 | 0 | 3449 ± 324 | 3371 ± 353 |
21 | −2 | 0 | 0 | 0 | 2413 ± 211 | 2416 ± 252 |
22 | 2 | 0 | 0 | 0 | 2506 ± 230 | 2492 ± 240 |
23 | 0 | −2 | 0 | 0 | 2302 ± 220 | 2535 ± 311 |
24 | 0 | 2 | 0 | 0 | 2242 ± 254 | 3115 ± 335 |
25 | 0 | 0 | −2 | 0 | 2375 ± 257 | 3154 ± 291 |
26 | 0 | 0 | 2 | 0 | 2352 ± 215 | 2476 ± 245 |
27 | 0 | 0 | 0 | −2 | 2782 ± 248 | 2894 ± 281 |
28 | 0 | 0 | 0 | 2 | 2414 ± 261 | 2291 ± 322 |
29 | 0 | 0 | 0 | 0 | 3379 ± 357 | 3371 ± 352 |
30 | 0 | 0 | 0 | 0 | 3425 ± 322 | 3371 ± 336 |
Source | Coefficient | Standard Error | t-Value | Prob > t |
---|---|---|---|---|
Intercept | 3371.8 | 74.6 | 45.2 | <0.0001 |
X1 | 18.958 | 37.3 | 0.5 | 0.6184 |
X2 | 145.1 | 37.3 | 3.9 | 0.0014 |
X3 | −169.5 | 37.3 | −4.6 | 0.0004 |
X4 | −150.6 | 37.3 | −4.0 | 0.0011 |
X1*X1 | −229.3 | 34.9 | −6.6 | <0.0001 |
X2*X1 | −100.2 | 45.7 | −2.2 | 0.0444 |
X2*X2 | −136.6 | 34.9 | −3.9 | 0.0014 |
X3*X1 | 79.1 | 45.7 | 1.7 | 0.1038 |
X3*X2 | 79.8 | 45.7 | 1.8 | 0.1009 |
X3*X3 | −139.1 | 34.9 | −4.0 | 0.0012 |
X4*X1 | 48.8 | 45.7 | 1.1 | 0.3019 |
X4*X2 | −20.7 | 45.7 | −0.5 | 0.6569 |
X4*X3 | 40.3 | 45.7 | 0.9 | 0.3912 |
X4*X4 | −194.7 | 34.9 | −5.6 | <0.0001 |
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Wei, Y.-H.; Wang, L.-C.; Chen, W.-C.; Chen, S.-Y. Production and Characterization of Fengycin by Indigenous Bacillus subtilis F29-3 Originating from a Potato Farm. Int. J. Mol. Sci. 2010, 11, 4526-4538. https://doi.org/10.3390/ijms11114526
Wei Y-H, Wang L-C, Chen W-C, Chen S-Y. Production and Characterization of Fengycin by Indigenous Bacillus subtilis F29-3 Originating from a Potato Farm. International Journal of Molecular Sciences. 2010; 11(11):4526-4538. https://doi.org/10.3390/ijms11114526
Chicago/Turabian StyleWei, Yu-Hong, Li-Chuan Wang, Wei-Chuan Chen, and Shan-Yu Chen. 2010. "Production and Characterization of Fengycin by Indigenous Bacillus subtilis F29-3 Originating from a Potato Farm" International Journal of Molecular Sciences 11, no. 11: 4526-4538. https://doi.org/10.3390/ijms11114526