Optimization of Moist and Oven-Dried Bacterial Cellulose Production for Functional Properties
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microbial Strain and Chemicals
2.2. Fermentation Method
2.3. Purification of Bacterial Cellulose (BC)
2.4. Processing of BC Membranes
2.5. Preliminary Study
2.5.1. Transmission Electron Microscopy (TEM)
2.5.2. Film Thickness and Uniformity
2.5.3. Film Weight and Yield
2.6. Optimization Study
2.6.1. Experimental Design and Target Optimal Levels for the Response Parameters
2.6.2. Fourier Transform Infrared Spectroscopy (FTIR)
2.6.3. Water Uptake Ability: Swelling Ratio and Moisture Content
2.6.4. Drug Release and Drug Half-Release Time
2.6.5. Mechanical Properties
2.6.6. Scanning Electron Microscopy (SEM)
2.6.7. Statistical Analysis, Response Surface Methodology (RSM), and Model Fitting
3. Results and Discussion
3.1. Preliminary Study
3.1.1. Transmission Electron Microscopy TEM
3.1.2. Film Thickness, Uniformity
3.1.3. Film Weight and Yield
3.2. Optimization Study
3.2.1. Fourier Transform Infrared Spectroscopy (FTIR)
3.2.2. Water Uptake Ability: Swelling Ratio and Moisture Content
3.2.3. Drug Release
3.2.4. Mechanical Properties
3.2.5. Scanning Electron Microscopy (SEM)
3.2.6. Optimization of Bacterial Cellulose (BC) by Response Surface Methodology (RSM)
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Model/Term | Linear | b0 | b1 | b2 | Square | b11 | b22 | Lack-of-Fit | R2 |
---|---|---|---|---|---|---|---|---|---|
Coefficient | 1.966 | 0.393 | −0.047 | −0.130 | 0.167 | 0.431 | 0.50 | ||
p-value | 0.000 | 0.000 | 0.000 | 0.354 | 0.054 | 0.143 | 0.053 | ||
Regression equation: | Y1 = 1.123 + 0.149 · X1 − 0.274 · X2 − 0.004 · X12 + 0.042 · X22 |
Independent Variables | Response—Dependent Variables | Desir | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
X1 Harvest (d) | X2 Inoculum Volume (mL) | X3 BC Type | Y1 Thickness * (mm) | Y2 Half-Swelling Time (h) | Y3 Drug Half-Release Time (h) | Y4 Tensile Strength σ (MPa) | Y5 Young’s Modulus E (MPa) | Y6 Fiber Diameter (nm) | ||||||||
exp | pred ** | exp | pred ** | exp | pred ** | exp | pred ** | exp | pred ** | exp | pred ** | |||||
1 | 6 | 1 | dry | 1.68 ± 0.16 bc | 1.62 | 1.25 ± 0.5 bc | 1.11 | 4.95 ± 0.77 de | 5.63 | 7.61 ± 0.21 ab | 7.88 | 128.92 ± 30.37 b | 118.19 | 51.34 ± 6.99 a | 50.18 | 0.40 |
2 | 18 | 1 | dry | 2.67 ± 0.67 ab | 2.81 | 1.92 ± 0.89 ab | 1.89 | 3.68 ± 0.32 e | 5.36 | 10.34 ± 3.69 a | 9.85 | 139.34 ± 22.35 b | 142.08 | 41.40 ± 3.87 de | 42.40 | 0.48 |
3 | 12 | 3 | dry | 2.09 ± 0.15 ab | 1.93 | 0.99 ± 0.38 c | 1.50 | 12.78 ± 3.45 a | 10.28 | 10.04 ± 1.90 a | 8.86 | 117.86 ± 28.18 b | 133.85 | 46.00 ± 7.61 bcd | 46.33 | 0.57 |
4 | 6 | 5 | dry | 1.34 ± 0.15 c | 1.05 | 1.22 ± 0.6 bc | 1.11 | 9.12 ± 1.60 b | 8.88 | 7.08 ± 2.78 abc | 7.88 | 143.99 ± 36.54 a | 160.75 | 47.11 ± 8.77 abc | 47.82 | 0.41 |
5 | 18 | 5 | dry | 2.28 ± 0.23 ab | 2.23 | 2.12 ± 0.84 a | 1.89 | 8.25 ± 1.61 bcd | 8.61 | 9.22 ± 3.33 a | 9.85 | 209.39 ± 23.85 c | 184.64 | 45.78 ± 6.05 bcd | 44.90 | 0.53 |
6 | 6 | 1 | moist | 1.68 ± 0.16 bc | 1.62 | 2.47 ± 0.20 a | 2.52 | 5.93 ± 0.58 bcde | 3.81 | 3.02 ± 0.64 d | 2.45 | 16.03 ± 2.97 c | 13.26 | 49.30 ± 4.18 ab | 50.54 | 0.29 |
7 | 18 | 1 | moist | 2.67 ± 0.67 ab | 2.81 | 2.53 ± 0.28 a | 2.54 | 3.77 ± 1.76 e | 3.54 | 4.64 ± 0.32 bcd | 4.42 | 26.38 ± 15.22 c | 37.15 | 43.67 ± 4.19 cde | 42.75 | 0.41 |
8 | 12 | 3 | moist | 2.09 ± 0.15 ab | 1.93 | 2.68 ± 0.18 a | 2.53 | 5.97 ± 2.25 bcde | 8.46 | 2.91 ± 0.83 d | 3.44 | 21.59 ± 11.90 c | 5.60 | 45.49 ± 2.64 bcde | 44.83 | 0.54 |
9 | 6 | 5 | moist | 1.34 ± 0.15 c | 1.05 | 2.49 ± 0.38 a | 2.52 | 5.38 ± 1.95 cde | 7.06 | 2.61 ± 0.38 d | 2.45 | 12.44 ± 0.73 c | 9.18 | 45.12 ± 6.03 bcde | 44.48 | 0.36 |
10 | 18 | 5 | moist | 2.28 ± 0.23 ab | 2.23 | 2.47 ± 0.08 a | 2.54 | 8.60 ± 2.81 bc | 6.79 | 4.00 ± 0.55 cd | 4.42 | 21.82 ± 2.47 c | 33.07 | 40.60 ± 4.99 e | 41.56 | 0.57 |
p-value *** | 0.992 | 0.824 | 0.853 | 0.897 | 0.971 | 0.912 |
Y1 Thickness (mm) |
Y2 Half-Swelling Time (h) |
Y3 Drug Half-Release Time (h) |
Y4 Tensile Strength σ (MPa) |
Y5 Young’s Modulus E (MPa) |
Y6 Fiber Diameter (nm) | Desirability | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
coef | p | coef | p | coef | p | coef | p | coef | p | coef | p | coef | p | ||
intercept | b0 | 1.926 *** | 0.000 | 2.013 *** | 0.000 | 9.370 *** | 0.000 | 6.149 *** | 0.000 | 69.720 *** | 0.000 | 45.580 *** | 0.000 | 0.553 *** | 0.000 |
linear | b1 | 0.591 *** | 0.002 | 0.203 * | 0.054 | −0.135 | 0.793 | 0.985 ** | 0.012 | 11.940 ** | 0.025 | −2.677 *** | 0.000 | 0.066 *** | 0.000 |
b2 | −0.289 *** | 0.000 | NA | NA | 1.627*** | 0.004 | NA | NA | 9.620 * | 0.066 | −0.889 * | 0.132 | 0.0339 *** | 0.000 | |
b3 (dry) | NA | NA | −0.514 *** | 0.000 | 0.911 * | 0.056 | 2.711 *** | 0.000 | 64.120 *** | 0.000 | 0.747 | 0.156 | 0.0216 *** | 0.000 | |
interaction | b12 | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | 1.215 ** | 0.044 | 0.0166 *** | 0.000 |
b13 | NA | NA | 0.191 * | 0.068 | NA | NA | NA | NA | NA | NA | NA | NA | −0.017 *** | 0.000 | |
b23 | NA | NA | NA | NA | NA | NA | NA | NA | 11.660 ** | 0.028 | 0.924 * | 0.118 | −0.0219 *** | 0.000 | |
square | b11 | NA | NA | NA | NA | −3.16 * | 0.010 | NA | NA | NA | NA | NA | NA | NA | NA |
b22 | NA | NA | NA | NA | NA | NA | NA | NA | 17.600 * | 0.129 | NA | NA | −0.123 *** | 0.000 | |
b33 | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | |
R2 | 0.70 | - | 0.61 | - | 0.47 | - | 0.73 | - | 0.90 | - | 0.59 | - | 0.995 | - | |
Lack-of-fit | - | 0.954 | - | 0.638 | - | 0.455 | - | 0.886 | - | 0.440 | - | 0.590 | - | - | |
The model | - | 0.000 | - | 0.000 | - | 0.003 | - | 0.000 | - | 0.000 | - | 0.000 | - | 0.000 |
Dry Bacterial Cellulose: | |
Y2 = 0.711 + 0.066·X1 | (4) |
Y3 = −4.540 + 2.086·X1+ 0.813·X2 − 0.088·X12 | (5) |
Y4 = 6.891 + 0.164·X1 | (6) |
Y5 = 117.600 − 15.700·X2 + 1.991·X1 + 4.390·X22 | (7) |
Y6 = 55.270 − 0.750·X1 − 1.197·X2 + 0.101·X1·X2 | (8) |
D = 0.232 + 0.173·X2 + 0.004·X1 - 0.031·X22 + 0.001·X2*X1 | (9) |
Moist bacterial cellulose: | |
Y2 = 2.504 + 0.002·X1 | (10) |
Y3 = −6.36 + 2.086·X1 + 0.813·X2 − 0.0879·X12 | (11) |
Y4 = 1.468 + 0.164·X1 | (12) |
Y5 = 24.300 + 1.991·X1 − 27.400·X2 + 4.390·X22 | (13) |
Y6 = 56.550 − 0.750·X1 − 2.121·X2 + 0.101·X1·X2 | (14) |
D = 0.055 + 0.010·X1 + 0.195·X2 + 0.001·X2·X1 − 0.031·X22 | (15) |
X1 = 16 d, X2 = 5 mL, and X3 = moist | X1 = 14 d, X2 = 4 mL, and X3 = dry | |||||||
---|---|---|---|---|---|---|---|---|
Resp. | pred. | 95% CI | 95% PI | exp. | pred. | 95% CI | 95% PI | exp. |
Y1 | 2.002 | (1.751; 2.252) | (1.134; 2.869) | 2.110 | 2.001 | (1.835; 2.168) | (1.154; 2.848) | 1.960 |
Y2 | 2.535 | (2.217; 2.852) | (1.474; 3.595) | 1.930 | 1.607 | (1.334; 1.880) | (0.559; 2.654) | 0.900 |
Y3 | 8.800 | (6.740; 10.870) | (3.270; 14.34) | 3.678 | 10.500 | (8.300; 12.700) | (4.91; 16.08) | 7.144 |
Y4 | 4.045 | (2.992; 5.098) | (0.230; 7.860) | 4.770 | 9.129 | (8.161; 10.097) | (5.337; 12.921) | 8.126 |
Y5 | 28.500 | (7.400; 49.600) | (6.300; 83.200) | 13.305 | 145.000 | (123.400; 166.600) | (90.100; 200.000) | 62.776 |
Y6 | 42.120 | (39.660; 44.580) | (35.850; 48.390) | 43.20 | 45.705 | (44.101; 47.310) | (39.721; 51.689) | 45.880 |
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Bodea, I.M.; Beteg, F.I.; Pop, C.R.; David, A.P.; Dudescu, M.C.; Vilău, C.; Stănilă, A.; Rotar, A.M.; Cătunescu, G.M. Optimization of Moist and Oven-Dried Bacterial Cellulose Production for Functional Properties. Polymers 2021, 13, 2088. https://doi.org/10.3390/polym13132088
Bodea IM, Beteg FI, Pop CR, David AP, Dudescu MC, Vilău C, Stănilă A, Rotar AM, Cătunescu GM. Optimization of Moist and Oven-Dried Bacterial Cellulose Production for Functional Properties. Polymers. 2021; 13(13):2088. https://doi.org/10.3390/polym13132088
Chicago/Turabian StyleBodea, Ioana M., Florin I. Beteg, Carmen R. Pop, Adriana P. David, Mircea Cristian Dudescu, Cristian Vilău, Andreea Stănilă, Ancuța M. Rotar, and Giorgiana M. Cătunescu. 2021. "Optimization of Moist and Oven-Dried Bacterial Cellulose Production for Functional Properties" Polymers 13, no. 13: 2088. https://doi.org/10.3390/polym13132088