Integrated Ultrasonication and Microbubble-Assisted Enzymatic Synthesis of Fructooligosaccharides from Brown Sugar
Abstract
:1. Introduction
2. Materials and Methods
2.1. Enzymes and Chemicals
2.2. Enzymatic Assay
2.3. Production of FOS Using Ultrasonication and Microbubbles
2.4. Optimisation of FOS Production
2.5. Characterisation of FOS
2.6. Statistical Analysis
3. Results and Discussion
3.1. Production of FOS
3.2. Optimisation of FOS Production
3.3. Production of FOS in a Large Scale
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Run | Variable | Enzyme Activity (U·mL–1) | Oligosaccharide Yields (mg·g–1) | |||||
---|---|---|---|---|---|---|---|---|
X1 (min) * | X2 (min) ** | X3 (h) *** | 1-Kestose | Nystose | 1F-β-Fructofuranosylnystose | Total FOS | ||
1 | 2.00 | 5.00 | 4.00 | 102.03 ± 1.88 | 379.80 ± 23.15 | 112.84 ± 1.55 | 10.30 ± 0.06 | 502.94 ± 35.46 |
2 | 15.00 | 5.00 | 4.00 | 97.75 ± 4.10 | 371.04 ± 34.06 | 108.94 ± 2.33 | 12.24 ± 0.15 | 492.22 ± 26.12 |
3 | 2.00 | 30.00 | 4.00 | 91.44 ± 2.30 | 351.19 ± 28.85 | 95.27 ± 1.69 | 8.31 ± 0.25 | 454.77 ± 44.59 |
4 | 15.00 | 30.00 | 4.00 | 95.13 ± 4.50 | 362.96 ± 49.73 | 103.95 ± 2.03 | 9.14 ± 0.59 | 476.05 ± 36.21 |
5 | 2.00 | 17.50 | 2.00 | 64.82 ± 0.98 | 311.30 ± 32.11 | 41.36 ± 0.96 | 2.09 ± 0.06 | 354.74 ± 25.89 |
6 | 15.00 | 17.50 | 2.00 | 70.88 ± 2.10 | 330.06 ± 44.53 | 52.42 ± 1.12 | 3.02 ± 0.12 | 385.50 ± 38.01 |
7 | 2.00 | 17.50 | 6.00 | 102.67 ± 2.71 | 341.32 ± 25.85 | 146.26 ± 3.54 | 18.65 ± 1.06 | 506.26 ± 46.04 |
8 | 15.00 | 17.50 | 6.00 | 112.22 ± 2.50 | 352.25 ± 25.40 | 148.59 ± 3.59 | 24.61 ± 2.09 | 525.45 ± 56.09 |
9 | 8.50 | 5.00 | 2.00 | 73.95 ± 2.59 | 313.25 ± 35.33 | 55.56 ± 0.58 | 2.96 ± 0.08 | 371.77 ± 34.56 |
10 | 8.50 | 30.00 | 2.00 | 70.87 ± 3.50 | 313.25 ± 27.56 | 50.43 ± 0.49 | 2.65 ± 0.11 | 366.32 ± 23.15 |
11 | 8.50 | 5.00 | 6.00 | 104.07 ± 3.15 | 358.53 ± 16.81 | 143.10 ± 4.32 | 17.28 ± 0.56 | 518.92 ± 35.11 |
12 | 8.50 | 30.00 | 6.00 | 105.04 ± 4.97 | 356.39 ± 40.74 | 149.94 ± 2.51 | 18.67 ± 0.92 | 525.00 ± 43.65 |
13 | 8.50 | 17.50 | 4.00 | 94.47 ± 1.39 | 367.94 ± 32.00 | 100.38 ± 2.09 | 8.32 ± 0.49 | 476.64 ± 35.28 |
14 | 8.50 | 17.50 | 4.00 | 99.41 ± 6.83 | 361.44 ± 42.75 | 120.14 ± 3.05 | 12.75 ± 0.46 | 494.33 ± 43.19 |
15 | 8.50 | 17.50 | 4.00 | 95.86 ± 2.11 | 354.56 ± 41.56 | 111.06 ± 1.54 | 10.62 ± 0.54 | 476.24 ± 54.56 |
16 | 8.50 | 17.50 | 4.00 | 95.60 ± 3.25 | 365.72 ± 26.35 | 109.90 ± 4.56 | 12.26 ± 0.83 | 487.89 ± 35.21 |
17 | 8.50 | 17.50 | 4.00 | 98.66 ± 2.17 | 359.31 ± 25.50 | 122.59 ± 3.97 | 12.89 ± 0.49 | 494.79 ± 43.54 |
Enzyme Activity | 1-Kestose | Nystose | 1F-β-Fructofuranosylnystose | Total FOS | ||||||
---|---|---|---|---|---|---|---|---|---|---|
F Value | p Value | F Value | p Value | F Value | p Value | F Value | p Value | F Value | p Value | |
Model | 24.57 | 0.0002 | 8.64 | 0.0048 | 75.78 | <0.0001 | 54.54 | <0.0001 | 42.45 | <0.0001 |
X1 * | 2.10 | 0.1904 | 0.985 | 0.354 | 2.13 | 0.168 | 3.18 | 0.0979 | 1.58 | 0.248 |
X2 ** | 2.18 | 0.1832 | 2.87 | 0.134 | 0.573 | 0.462 | 0.545 | 0.473 | 3.78 | 0.0931 |
X3 *** | 191.53 | <0.0001 | 27.92 | 0.0011 | 224.63 | <0.0001 | 159.89 | <0.0001 | 310.96 | <0.0001 |
X1X2 | 1.18 | 0.3135 | 2.03 | 0.197 | 1.91 | 0.210 | ||||
X1X3 | 0.225 | 0.6494 | 0.173 | 0.690 | 1.76 | 0.226 | ||||
X2X3 | 0.304 | 0.5984 | 0.0128 | 0.913 | 0.247 | 0.634 | ||||
X12 | 0.0859 | 0.7780 | 0.230 | 0.646 | 1.66 | 0.239 | ||||
X22 | 0.0307 | 0.8660 | 0.694 | 0.432 | 0.00004 | 0.995 | ||||
X32 | 23.34 | 0.0019 | 43.55 | 0.0003 | 58.68 | 0.0001 | ||||
Lack of Fit | 5.61 | 0.0645 | 6.06 | 0.0572 | 1.27 | 0.438 | 0.981 | 0.553 | 4.34 | 0.0951 |
Response | Final Equation in Term of Actual Factors | P * | LOF | R2 | Adj. R2 | AP | CV |
---|---|---|---|---|---|---|---|
Enzyme activity | 35.10–0.20 × X1–0.59 × X2 + 24.95 × X3–0.012 × X12 + 0.002 × X22–2.16 × X32 + 0.024 × X1 × X2 + 0.07 × X1 × X3 + 0.04 × X2 × X3 | 0.0002 | 0.0645 | 0.9693 | 0.9299 | 14.480 | 3.96 |
1-Kestose | 226.07–1.22 × X1–1.92 × X2 + 70.97 × X3 + 0.01 × X12 + 0.02 × X22–7.57 × X32 + 0.08 × X1 ×X2–0.15 × X1 ×X3–0.02 × X2 × X3 | 0.0048 | 0.0572 | 0.9175 | 0.8113 | 9.140 | 2.69 |
Nystose | 12141 + 0.77 × X1–0.21 × X2 + 25.63 × X3 | <0.0001 | 0.4385 | 0.9459 | 0.9334 | 23.978 | 9.17 |
1F-β-Fructofuranosyl-nystose | –7.02 + 0.19 × X1–0.04 × X2 + 4.28 × X3 | <0.0001 | 0.5529 | 0.9264 | 0.9094 | 21.032 | 17.44 |
Total FOS | 165.75 + 4.35 × X1–1.94 × X2 + 125.68 × X3–0.17 × X12 + 0.0002 × X22–10.82 × X32 + 0.10 × X1 × X2–0.59 × X1 × X3 + 0.12 × X2 × X3 | <0.0001 | 0.0951 | 0.9820 | 0.9589 | 19.098 | 2.49 |
Factors | Pectinex Ultra SP-L Activity (U·mL−1) | Fructooligosaccharide Yields (mg·g−1) | |||
---|---|---|---|---|---|
1-Kestose | Nystose | 1F-β-Fructofurano-sylnystose | Total FOS | ||
Brown sugar using a combination of ultrasonication and microbubble | 102.51 ± 4.69 | 368.02 ± 20.22 | 110.53 ± 8.81 | 16.35 ± 1.49 | 494.89 ± 19.98 |
White sugar using a combination of ultrasonication and microbubbles | 107.93 ± 5.26 | 415.02 ± 19.54 | 147.50 ± 9.84 | 17.49 ± 2.94 | 580.01 ± 21.35 |
White sugar using only ultrasonication | 90.05 ± 3.54 | 399.48 ± 21.33 | 103.83 ± 12.33 | 10.23 ± 1.66 | 515.53 ± 20.43 |
Substrate | Enzyme | Condition | Reaction Time (h) | FOS Yield (g·g−1 Substrate) | Reference |
---|---|---|---|---|---|
Brown sugar | Pectinex Ultra SP-L | Ultrasonication and microbubble, 55 °C, 700 g·L−1 substrate | 4 | 0.45 | This study |
White sugar | Pectinex Ultra SP-L | Ultrasonication and microbubble, 55 °C, 700 g·L−1 substrate | 5.67 | 0.58 | This study |
White sugar | GAPfopA_V1 (engineered β-fructofuronosidase) | Water bath, 62 °C, 120 rpm, 600 g·L−1 substrate | 5. 6 | 0.55 | [43] |
White sugar | Pectinex Ultra SP-L | Ultra-high pressure at 300 MPa, 30 °C, 600 g·L−1 substrate | 0.25 | 0.57 | [5] |
White sugar and Jerusalem artichoke | Inulinase | Water bath, 55 °C, 59.4 g·L−1 inulin and 598.7 g·L−1 refine sugar | 9 | 0.67 | [38] |
White sugar | Pectinex Ultra SP-L | Water bath, 60 °C, 400 g·L−1 substrate | 16 | 0.62 | [39] |
White sugar | Pectinex Ultra SP-L | Water bath, 55 °C, 500 g·L−1 substrate | 10 | 0.60 | [40] |
White sugar | Pectinex Ultra SP-L | Water bath, 50 °C, 536.2 g·L−1 substrate | 6 | 0.59 | [41] |
White sugar | Culture fluid of Aspergillus oryzae CFR202 | Water bath, 55 °C, 64.55 g·L−1 substrate | 12 | 0.58 | [42] |
Method | Reaction Time (h) | Fructooligosaccharide | Reference | |||||||
---|---|---|---|---|---|---|---|---|---|---|
Concentration (g·L−1) | Productivity (g·L−1·h−1) | |||||||||
GF2 * | GF3 | GF4 | Total FOS | GF2 | GF3 | GF4 | Total FOS | |||
Ultrasonic and microbubble methods | 4 | 264.1 | 47.8 | 4.84 | 316.8 | 66.0 | 11.96 | 1.21 | 79.2 | This study |
Ultra-high pressure (UHP) method | 0.25 | 189.72 | 110.88 | 28.2 | 328.8 | 758.88 | 443.52 | 112.8 | 1315.2 | [5] |
Conventional methods | 5.6 | 220.8 | 321.6 | 60.0 | 330 | 39.4 | 57.4 | 10.7 | 58.9 | [43] |
9 | 156.98 | 144.10 | 84.63 | 404.0 | 17.44 | 16.01 | 9.40 | 44.90 | [38] | |
10 | 155 | 130 | 15.0 | 300 | 15.5 | 13.0 | 1.50 | 30.0 | [40] | |
6 | 212.33 | 106.7 | 8.58 | 327.6 | 35.4 | 17.78 | 1.43 | 54.6 | [41] | |
12 | 174.2 | 96.6 | 26.8 | 297.7 | 14.5 | 8.05 | 2.24 | 24.8 | [42] |
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Chaisuwan, W.; Manassa, A.; Phimolsiripol, Y.; Jantanasakulwong, K.; Chaiyaso, T.; Pathom-aree, W.; You, S.; Seesuriyachan, P. Integrated Ultrasonication and Microbubble-Assisted Enzymatic Synthesis of Fructooligosaccharides from Brown Sugar. Foods 2020, 9, 1833. https://doi.org/10.3390/foods9121833
Chaisuwan W, Manassa A, Phimolsiripol Y, Jantanasakulwong K, Chaiyaso T, Pathom-aree W, You S, Seesuriyachan P. Integrated Ultrasonication and Microbubble-Assisted Enzymatic Synthesis of Fructooligosaccharides from Brown Sugar. Foods. 2020; 9(12):1833. https://doi.org/10.3390/foods9121833
Chicago/Turabian StyleChaisuwan, Worraprat, Apisit Manassa, Yuthana Phimolsiripol, Kittisak Jantanasakulwong, Thanongsak Chaiyaso, Wasu Pathom-aree, SangGuan You, and Phisit Seesuriyachan. 2020. "Integrated Ultrasonication and Microbubble-Assisted Enzymatic Synthesis of Fructooligosaccharides from Brown Sugar" Foods 9, no. 12: 1833. https://doi.org/10.3390/foods9121833