Papain Covalently Immobilized on Chitosan–Clay Nanocomposite Films: Application in Synthetic and Real White Wine
Abstract
:1. Introduction
2. Materials and Methods
2.1. Enzyme, Chemicals and Wines
2.2. Preparation of CS–Clay Nanocomposite Films by Solvent Casting
2.3. Physical Characterization of CS–Clay Nanocomposite Films
2.4. Papain Immobilization on CS–Clay Nanocomposite Films
2.5. Proteolytic Activity Assay
2.6. Kinetic Characterization of Immobilized Papain
2.7. Wine Stabilization Treatment in the Batch-Scale Stirred Reactor
2.8. Wine Protein Content Determination
2.9. Heat Test
2.10. Statistical Analysis
3. Results and Discussion
3.1. Physical Properties of CS–Clay Nanocomposite Films
3.2. Kinetic Properties of Papain Immobilized on CS–Clay Nanocomposite Films
3.3. Wine Stabilization Treatment in Batch-Scale Stirred Reactor
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Manzoni | Sauvignon Blanc | |
---|---|---|
pH | 3.37 ± 0.01 | 3.34 ± 0.01 |
Total acidity (g/L tartaric acid) | 6.00 ± 0.05 | 6.00 ± 0.05 |
Alcohol level (% v/v) | 11.2 ± 0.3 | 13.2 ± 0.2 |
Free SO2 (mg/L) | 6 ± 1 | 10 ± 1 |
Total SO2 (mg/L) | 30 ± 2 | 48 ± 3 |
Total phenols (mg/L catechin) | 217 ± 1 | 219 ± 4 |
Total protein (mg/L) | 465 ± 43 | 120 ± 7 |
ΔNTU Index0 | 600 ± 11 | 116 ± 8 |
Clay-Free | OPT | SMP | |||||
---|---|---|---|---|---|---|---|
20 | 30 | 50 | 20 | 30 | 50 | ||
TIHB (°C) | 121 ± 0.5 | 118 ± 0.4 | 124 ± 0.7 | 111 ± 0.6 | 118 ± 0.4 | 115 ± 0.4 | 105 ± 0.3 |
Td (°C) | 287 ± 1.2 | 274 ± 1.3 | 272 ± 0.9 | 260 ± 1.0 | 279 ± 1.1 | 280 ± 1.4 | 284 ± 1.5 |
ΔHmI (J/g) | 259 ± 1.5 | 259 ± 1.4 | 238 ± 1.2 | 142 ± 0.9 | 274 ± 1.1 | 296 ± 1.3 | 335 ± 2.0 |
E (MPa) | σmax (MPa) | εmax | |
---|---|---|---|
Clay-Free | 579 ± 23 | 16 ± 1 | 0.30 ± 0.01 |
OPT-20 | 738 ± 20 | 21± 6 | 0.22 ± 0.08 |
OPT-30 | 653 ± 71 | 23 ± 4 | 0.13 ± 0.05 |
OPT-50 | 1670 ± 202 | 36 ± 7 | 0.06 ± 0.03 |
SMP-20 | 580 ± 55 | 24 ± 4 | 0.28 ± 0.05 |
SMP-30 | 528 ± 4 | 22 ± 3 | 0.12 ± 0.03 |
SMP-50 | 1582 ± 210 | 19 ± 4 | 0.04 ± 0.01 |
Sample | IY (%) | Vmax (mIU mg−1PI) | KM (μM) | kcat (min−1) | Ka (min−1 μM−1) | R2 |
---|---|---|---|---|---|---|
Clay-free | 45 ± 6 b,c | 6.9 ± 0.2 e | 88 ± 9 a | 3183 ± 0 e | 36 ± 1 d | 0.99 |
OPT-20 | 41 ± 4 c,d | 15.3 ± 0.9 c | 75 ± 16 a,b | 6535 ± 1 c | 87 ± 4 c | 0.97 |
OPT-30 | 26 ± 5 e | 21.4 ± 1.0 b | 106 ± 23 a | 12507 ± 1 b | 118 ± 6 b | 0.98 |
OPT-50 | 29 ± 2 de | 38.9 ± 2.0 a | 102 ± 14 a | 20417 ± 2 a | 199 ± 3 a | 0.99 |
OPT-70 | 55 ± 3 b | 11.6 ± 0.5 d | 50 ± 8 b,c | 4225 ± 0 d | 85 ± 3 c | 0.98 |
SMP-20 | 54 ± 7 a,b,c | 2.6 ± 0.1 f | 34 ± 8 c | 1017 ± 0 h | 30 ± 2 d,e | 0.97 |
SMP-30 | 41 ± 5 c,d | 3.5 ± 0.2 f | 46 ± 11 b,c | 642 ± 0 i | 14 ± 1 f | 0.97 |
SMP-50 | 56 ± 2 b | 4.5 ± 0.1 f | 49 ± 2 b,c | 1221 ± 0 g | 25 ± 0 e | 1.00 |
SMP-70 | 60 ± 4 a | 7.3 ±0.1 e | 29 ± 3 c | 2503 ± 0 f | 86 ± 1 c | 1.00 |
Treatment | Net Haze after Heat Test | Residual Protein Content | ||
---|---|---|---|---|
ΔNTU | Reduction (TRY, %) | mg BSAeq/L | Reduction (%) | |
Manzoni | ||||
Untreated wine | 601 ± 11 a | - | 465 ± 63 a | - |
OPT-50 | 102 ± 41 b | 83 | 124 ± 35 b | 73 |
Sauvignon Blanc | ||||
Untreated wine | 116 ± 8 a | - | 124 ± 1 a | - |
OPT-50 | 80 ± 1 b | 31 | 109 ± 1 b | 12 |
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Benucci, I.; Lombardelli, C.; Cacciotti, I.; Esti, M. Papain Covalently Immobilized on Chitosan–Clay Nanocomposite Films: Application in Synthetic and Real White Wine. Nanomaterials 2020, 10, 1622. https://doi.org/10.3390/nano10091622
Benucci I, Lombardelli C, Cacciotti I, Esti M. Papain Covalently Immobilized on Chitosan–Clay Nanocomposite Films: Application in Synthetic and Real White Wine. Nanomaterials. 2020; 10(9):1622. https://doi.org/10.3390/nano10091622
Chicago/Turabian StyleBenucci, Ilaria, Claudio Lombardelli, Ilaria Cacciotti, and Marco Esti. 2020. "Papain Covalently Immobilized on Chitosan–Clay Nanocomposite Films: Application in Synthetic and Real White Wine" Nanomaterials 10, no. 9: 1622. https://doi.org/10.3390/nano10091622