The Preparation and Physiochemical Characterization of Tenebrio molitor Chitin Using Alcalase
Abstract
:1. Introduction
2. Results and Discussions
2.1. Protease Screening for Protein Removal from Mealworms
2.2. Establishment of Optimal Alcalase Hydrolysis Conditions for Mealworm Protein Removal by RSM
2.3. FT-IR Spectroscopy of Chitin and Chitosan
2.4. Thermodynamic Properties of Chitin and Chitosan Using DSC
2.5. Degree of Deacetylation (DDA) and Solubility of Chitosan
3. Materials and Methods
3.1. Materials
3.2. Mealworm Protein Hydrolysis and Deacetylation
3.3. Determination of Degree of Deproteinization (DP)
3.4. Optimization of Protein Removal Conditions by Alcalase by Response Surface Methodology (RSM)
3.5. α-Amino Nitrogen (AN) and Crude Protein Analysis
3.6. Solubility (%) of Chitosan Derived from Mealworm
3.7. FT-IR Spectroscopy of Chitin and Chitosan
3.8. Thermal Characterization of Chitin and Chitosan by Differential Scanning Calorimetry (DSC)
3.9. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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RSM Result | Actual Result | ||||||
---|---|---|---|---|---|---|---|
Substrate (g) | Enzyme (μL) | Time (h) | Amino-Nitrogen (mg/g of Sample) | Substrate (g) | Enzyme (μL) | Time (h) | Amino-Nitrogen (mg/g of Sample) |
7.5 | 80 | 24 | 163.17 | 7.5 | 80 | 24 | 162.99 |
Sample | Exotherm (°C) | Endotherm (°C) | ||||||
---|---|---|---|---|---|---|---|---|
Ts1 (°C) | Tmax1 (°C) | Te1 (°C) | ΔH1 (J/g) | Ts2 (°C) | Tmax2 (°C) | Te2 (°C) | ΔH2 (J/g) | |
commercial chitin | 140.77 | 143.29 | 157.37 | −116.6 | 349.90 | 385.40 | 397.15 | 17.848 |
mealworm chitin | 132.82 | 153.76 | 179.76 | −91.08 | 325.22 | 359.82 | 394.48 | 24.075 |
commercial chitosan | 140.77 | 143.65 | 165.68 | −183.1 | 281.20 | 306.47 | 331.38 | 104.7 |
mealworm chitosan | 141.13 | 147.63 | 197.81 | −179.9 | 301.06 | 337.52 | 371.82 | 181.7 |
Mealworm Chitosan | Commercial Chitosan | |
---|---|---|
Degree of deacetylation (%) | 37.34 ± 4.35 | 57.58 ± 7.88 |
Solubility (%) | 45.94 ± 15.33 | 61.34 ± 15.96 |
Run Order | Coded Variables | Real Variables | Amino-Nitrogen (mg/g of Sample) | ||||
---|---|---|---|---|---|---|---|
X | Y | Z | X Substrate Concentration (%) | Y Enzyme Concentration (%) | Z Time (h) | ||
1 | −1.00000 | −1.00000 | −1.00000 | 5 | 50 | 6 | 83.46 ± 0.33 |
2 | 0.00000 | −1.68179 | 0.00000 | 7.5 | 25 | 8 | 94.93 ± 4.35 |
3 | 1.0000 | −1.0000 | −1.00000 | 10 | 50 | 6 | 83.49 ± 0.06 |
4 | 1.68179 | 0.00000 | 0.00000 | 12.5 | 80 | 8 | 122.03 ± 0.94 |
5 | 1.00000 | 1.00000 | −1.00000 | 10 | 100 | 6 | 99.45 ± 3.48 |
6 | 0.00000 | 0.00000 | 1.68179 | 7.5 | 80 | 24 | 162.99 ± 4.68 |
7 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 118.61 ± 0.63 |
8 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 119.66 ± 1.60 |
9 | 1.00000 | 1.00000 | 1.00000 | 10 | 100 | 12 | 148.87 ± 3.34 |
10 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 122.48 ± 1.77 |
11 | −1.00000 | −1.00000 | −1.00000 | 5 | 50 | 6 | 78.42 ± 0.78 |
12 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 121.15 ± 3.01 |
13 | −1.68179 | 0.00000 | 0.00000 | 2.5 | 80 | 8 | 132.79 ± 1.38 |
14 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 116.00 ± 2.08 |
15 | −1.00000 | 1.00000 | 1.00000 | 5 | 100 | 12 | 157.99 ± 0.92 |
16 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 120.07 ± 0.20 |
17 | 0.00000 | 0.00000 | −1.68179 | 7.5 | 80 | 4 | 65.57 ± 1.74 |
18 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 121.79 ± 3.02 |
19 | −1.00000 | −1.00000 | 1.00000 | 5 | 50 | 12 | 127.91 ± 1.14 |
20 | 1.00000 | −1.00000 | −1.00000 | 10 | 50 | 6 | 83.45 ± 5.49 |
21 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 122.96 ± 2.51 |
22 | 0.00000 | −1.68179 | 0.00000 | 7.5 | 25 | 8 | 97.14 ± 1.94 |
23 | 1.68179 | 0.00000 | 0.00000 | 12.5 | 80 | 8 | 130.52 ± 1.02 |
24 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 129.01 ± 0.79 |
25 | 0.00000 | 0.00000 | 1.68179 | 7.5 | 80 | 24 | 162.01 ± 2.17 |
26 | 0.00000 | 0.00000 | −1.68179 | 7.5 | 80 | 4 | 67.89 ± 3.08 |
27 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 120.45 ± 1.74 |
28 | −1.00000 | 1.00000 | −1.00000 | 5 | 100 | 6 | 94.96 ± 0.41 |
29 | 1.00000 | 1.00000 | −1.00000 | 10 | 100 | 6 | 99.25 ± 0.57 |
30 | 0.00000 | 1.68179 | 0.00000 | 7.5 | 130 | 8 | 136.45 ± 4.17 |
31 | −1.00000 | −1.00000 | 1.00000 | 5 | 50 | 12 | 130.30 ± 2.66 |
32 | −1.68179 | 0.00000 | 0.00000 | 2.5 | 80 | 8 | 127.57 ± 1.31 |
33 | 1.00000 | 1.00000 | 1.00000 | 10 | 100 | 12 | 155.39 ± 1.99 |
34 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 119.53 ± 2.70 |
35 | 0.00000 | 1.68179 | 0.00000 | 7.5 | 130 | 8 | 139.70 ± 2.11 |
36 | 1.00000 | −1.00000 | 1.00000 | 10 | 50 | 12 | 132.45 ± 4.34 |
37 | 1.00000 | −1.00000 | 1.00000 | 10 | 50 | 12 | 131.45 ± 2.71 |
38 | −1.00000 | 1.00000 | −1.00000 | 5 | 100 | 6 | 94.92 ± 2.38 |
39 | 0.00000 | 0.00000 | 0.00000 | 7.5 | 80 | 8 | 125.42 ± 1.43 |
40 | −1.00000 | 1.00000 | 1.00000 | 5 | 100 | 12 | 159.73 ± 1.60 |
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Kim, H.; Kim, H.; Ahn, Y.; Hong, K.-B.; Kim, I.-W.; Choi, R.-Y.; Suh, H.J.; Han, S.H. The Preparation and Physiochemical Characterization of Tenebrio molitor Chitin Using Alcalase. Molecules 2023, 28, 3254. https://doi.org/10.3390/molecules28073254
Kim H, Kim H, Ahn Y, Hong K-B, Kim I-W, Choi R-Y, Suh HJ, Han SH. The Preparation and Physiochemical Characterization of Tenebrio molitor Chitin Using Alcalase. Molecules. 2023; 28(7):3254. https://doi.org/10.3390/molecules28073254
Chicago/Turabian StyleKim, Hyemi, Hyeongyeong Kim, Yejin Ahn, Ki-Bae Hong, In-Woo Kim, Ra-Yeong Choi, Hyung Joo Suh, and Sung Hee Han. 2023. "The Preparation and Physiochemical Characterization of Tenebrio molitor Chitin Using Alcalase" Molecules 28, no. 7: 3254. https://doi.org/10.3390/molecules28073254