Electrospinning Microencapsulation of Lactobacillus fermentum K73 Using Gelatin as the Main Component of a Food-Grade Matrix
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Bacterial Strain and Culture Conditions
2.3. Preparation of Carrier Material
2.3.1. Mixture Experimental Design to Select the Ratio of Wall Materials
2.3.2. Mixture Characterization
2.4. Improvement in Probiotic Viability during the Electrospinning Process through Box–Behnken Design
2.4.1. Electrospinning Process
2.4.2. Cell Count
2.4.3. Scanning Electron Microscopy (SEM)
2.5. Survival Percentage of Encapsulated L. fermentum K73 during Simulated Gastrointestinal Conditions
2.6. Statistical Analysis
3. Results
3.1. Characterization of Polymeric Solutions of the Mixture Experimental Design
3.2. Formulation of Carrier Material
3.3. Cell Counting Process: Release of Probiotics from Electrospun Fibers
3.4. Improvement of Encapsulation Yield in the Electrospinning Process
3.5. Morphological Characteristics
3.6. Tolerance of Encapsulated and Non-Encapsulated Probiotic to Simulated Gastrointestinal Conditions
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Mixture | Ratio | Surface Tension (mN m−1) | Viscosity (mPa s−1) | Conductivity (mS cm−1) | Ph | |
---|---|---|---|---|---|---|
Gelatin | Culture | |||||
1 | 0.20 | 0.80 | 42.9 ± 0.03 a | 4.80 ± 0.28 d | 4.88 ± 0.25 a | 4.10 ± 0.06 a |
2 | 0.35 | 0.65 | 39.3 ± 0.025 b | 7.90 ± 0.70 d | 4.10 ± 0.17 b | 4.20 ± 0.00 a |
3 | 0.40 | 0.60 | 40.8 ± 1.76 ab | 19.55 ± 0.49 c | 3.76 ± 0.08 b | 4.40 ± 0.11 a |
4 | 0.50 | 0.50 | 42.0 ± 0.62 ab | 23.90 ± 0.56 c | 1.95 ± 0.03 c | 4.40 ± 0.05 a |
5 | 0.60 | 0.40 | 42.6 ± 0.98 a | 33.95 ± 2.19 b | 1.73 ± 0.04 c | 4.40 ± 0.04 a |
6 | 0.65 | 0.35 | 42.7 ± 1.08 a | 34.10 ± 1.6 b | 1.58 ± 0.00 c | 4.50 ± 0.19 a |
7 | 0.80 | 0.20 | 42.5 ± 0.82 a | 48.58 ± 1.38 a | 1.92 ± 0.07 c | 4.90 ± 0.02 a |
Correlation | Pearson Correlation Coefficient | p-Value |
---|---|---|
A/B and pH | 0.937 | 0.002 |
A/B and viscosity | 0.915 | 0.004 |
B/A and conductivity | 0.916 | 0.004 |
B/A and pH | −0.764 | 0.045 |
B/A and viscosity | −0.858 | 0.013 |
B/A and bacterial cycles change under bile salts | −0.830 | 0.021 |
Conductivity and bacterial cycles change under bile salts | −0.953 | 0.001 |
Run | Factor wt% (Wall Material) | Response Variable: Bacterial Cycles Change log (CFU/mL) | |||
---|---|---|---|---|---|
Gelatin [A] | Culture [B] | After Mixture | Gastric pH | Bile Salt | |
1 | 0.40 | 0.60 | 0.04 | 0.09 | −1.87 |
2 | 0.65 | 0.35 | 0.19 | −0.16 | −1.25 |
3 | 0.20 | 0.80 | −0.07 | −3.18 | −2.48 |
4 | 0.60 | 0.40 | 0.10 | −0.56 | −1.11 |
5 | 0.50 | 0.50 | 0.05 | 0.01 | −1.22 |
6 | 0.80 | 0.20 | −0.05 | −2.05 | −1.39 |
7 | 0.20 | 0.80 | −0.06 | −3.20 | −1.98 |
8 | 0.80 | 0.20 | −0.08 | −3.15 | −1.70 |
9 | 0.35 | 0.65 | 0.15 | −1.59 | −1.59 |
10 | 0.80 | 0.20 | −0.08 | −2.01 | −1.55 |
p-value | 0.0008 | 0.0005 | 0.0080 |
Source | Change in Bacterial Cycles | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
After Mixture | Gastric pH | Bile Salts | |||||||||||||
SS * | Df ** | Mean Square | F-Value | p-Value | SS * | Df ** | Mean Square | F-Value | p-Value | SS * | Df ** | Mean Square | F-Value | p-Value | |
Model | 0.09 | 4 | 0.02 | 34.5 | 0.0008 | 14.6 | 2 | 7.29 | 27.6 | 0.0005 | 1.39 | 2 | 0.69 | 10.4 | 0.0080 |
Linear mixture | 6.19 × 10−4 | 1 | 6.19 × 10−4 | 0.95 | 0.3740 | 0.280 | 1 | 0.280 | 1.05 | 0.3399 | 0.59 | 1 | 0.59 | 8.89 | 0.0204 |
AB | 9.07 × 10−3 | 1 | 9.07 × 10−3 | 13.9 | 0.0135 | 14.3 | 1 | 14.3 | 54.1 | 0.0002 | 0.80 | 1 | 0.80 | 11.90 | 0.0107 |
AB(A-B) | 2.25 × 10−3 | 1 | 2.25 × 10−3 | 3.46 | 0.1220 | ||||||||||
AB(A-B)2 | 0.02 | 1 | 0.023 | 34.7 | 0.0020 | ||||||||||
Residual | 3.25 × 10−3 | 5 | 6.50 × 10−4 | 1.85 | 7 | 0.260 | 0.47 | 7 | 0.07 | ||||||
Lack of fit | 2.60 × 10−3 | 2 | 1.30 × 10−3 | 6.00 | 0.0894 | 1.01 | 4 | 0.250 | 0.91 | 0.5543 | 0.29 | 4 | 0.07 | 1.28 | 0.4376 |
Pure error | 6.50 × 10−4 | 3 | 2.17 × 10−4 | 0.84 | 3 | 0.280 | 0.17 | 3 | 0.06 | ||||||
Corr. Total | 0.09 | 9 | 16.43 | 9 | 1.86 | 9 | |||||||||
R2 | 0.96 | 0.88 | 0.74 | ||||||||||||
Equation | N (log CFU/mL) = −0.045358 ∗ A − 0.040809 ∗ B + 1.73313 × 10−3 ∗ A ∗ B + 1.39291 × 10−6 ∗ B ∗ A ∗ (A − B) + 2.54879 × 10−7 ∗ A∗ B ∗ (A − B)2 | N (log CFU/mL Log) = −0.070443 ∗ A − 0.084910 ∗ B + 3.09535 × 10−3 ∗ A ∗ B | N (log CFU/mL) = 0.024460 ∗ A − 0.036966 ∗ B + 7.29884 × 10−4 ∗ A ∗ B |
Factors | Response Variable Encapsulation Yield and Survival (%) | |||
---|---|---|---|---|
Run | Bacteria Culture (g/L) | Soy Lecithin (%wt/v) | Collector Distance (cm) | |
1 | 150 | 5.0 | 9.0 | 65 |
2 | 200 | 2.5 | 9.0 | 67 |
3 | 150 | 2.5 | 8.0 | 66 |
4 | 100 | 2.5 | 7.0 | 73 |
5 | 200 | 2.5 | 7.0 | 87 |
6 | 150 | 2.5 | 8.0 | 69 |
7 | 100 | 5.0 | 8.0 | 65 |
8 | 150 | 2.5 | 8.0 | 70 |
9 | 100 | 0.0 | 8.0 | 75 |
10 | 100 | 2.5 | 9.0 | 71 |
11 | 150 | 2.5 | 8.0 | 68 |
12 | 150 | 5.0 | 7.0 | 63 |
13 | 200 | 5.0 | 8.0 | 63 |
14 | 150 | 0.0 | 9.0 | 86 |
15 | 150 | 2.5 | 8.0 | 61 |
Sum of Squares | DF * | Mean Square | F-Value | p-Value | |
---|---|---|---|---|---|
Model | 752.71 | 9 | 83.63 | 4.98 | 0.0459 |
[A]-Bacteria culture | 76.80 | 1 | 76.80 | 4.57 | 0.0855 |
[B]-Soy lecithin | 450.67 | 1 | 450.67 | 26.83 | 0.0035 |
[C]-Collector distance | 76.80 | 1 | 76.80 | 4.57 | 0.0855 |
AB | 78.11 | 1 | 78.11 | 4.65 | 0.0835 |
AC | 72.25 | 1 | 72.25 | 4.30 | 0.0928 |
BC | 24.11 | 1 | 24.11 | 1.44 | 0.2846 |
A2 | 17.14 | 1 | 17.14 | 1.02 | 0.3587 |
B2 | 70.08 | 1 | 70.08 | 4.17 | 0.0965 |
C2 | 89.05 | 1 | 89.05 | 5.30 | 0.0695 |
Residual | 83.97 | 5 | 16.79 | ||
Lack of Fit | 37.50 | 1 | 37.50 | 3.23 | 0.1468 |
Pure Error | 46.47 | 4 | 11.62 | ||
R2 | 0.899 |
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Serrano-Delgado, A.; Quintanilla-Carvajal, M.X. Electrospinning Microencapsulation of Lactobacillus fermentum K73 Using Gelatin as the Main Component of a Food-Grade Matrix. Microorganisms 2023, 11, 2682. https://doi.org/10.3390/microorganisms11112682
Serrano-Delgado A, Quintanilla-Carvajal MX. Electrospinning Microencapsulation of Lactobacillus fermentum K73 Using Gelatin as the Main Component of a Food-Grade Matrix. Microorganisms. 2023; 11(11):2682. https://doi.org/10.3390/microorganisms11112682
Chicago/Turabian StyleSerrano-Delgado, Arjana, and María Ximena Quintanilla-Carvajal. 2023. "Electrospinning Microencapsulation of Lactobacillus fermentum K73 Using Gelatin as the Main Component of a Food-Grade Matrix" Microorganisms 11, no. 11: 2682. https://doi.org/10.3390/microorganisms11112682