Structural Properties of Cruciferin and Napin of Brassica napus (Canola) Show Distinct Responses to Changes in pH and Temperature
Abstract
:1. Introduction
2. Results
2.1. Proteins of B. napus Meal and Their Solubility Behaviour with pH Change
2.2. Obtaining Cruciferin
2.3. Obtaining Napin
2.4. Secondary Structure Features of Cruciferin and Napin and Their Changes Due to Medium pH
2.5. Tertiary Structure Details of Cruciferin and Napin and Their Changes with Medium pH
2.6. Thermal Stability of Cruciferin and Napin Structure at Different pHs
3. Discussion
3.1. Obtaining Cruciferin and Napin
3.2. Effect of pH and Temperature on Cruciferin
3.3. Effect of pH and Temperature on Napin
3.4. Relevance to Seed Protein Utilization
4. Materials and Methods
4.1. Seeds
4.2. Meal and Meal Protein
4.2.1. Preparation of Meal
4.2.2. Types of Protein Soluble at Different pHs
4.3. Preparation of Purified Cruciferin
4.3.1. Extracting Meal Protein
4.3.2. Separation and Purification of Cruciferin
4.4. Isolation and Purification of Napin at Low pH
4.5. N-Based Protein Content
4.6. Sodium Dodecyl Sulfite Polyacrylamide Gel Electrophoresis (SDS-PAGE)
4.7. Native Polyacrylamide Gel Electrophoresis (Native-PAGE)
4.8. FT-IR Spectroscopy
4.9. Circular Dichroism (CD) Spectroscopy
4.10. Fluorescence Spectroscopy
4.10.1. Intrinsic Fluorescence
4.10.2. Surface Hydrophobicity
4.11. Differential Scanning Calorimetry (DSC)
4.12. Experimental Design and Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Protein | pH | α-Helix | β-Sheet | β-Turn | Random | Surface Hydrophobicity |
---|---|---|---|---|---|---|
(%) | (%) | (%) | (%) | (S0) 2 | ||
Cruciferin | 3 | 10.7 ±1.0 a | 25.4 ± 3.3 a | 26.0 ± 0.7 a | 38.0 ± 3.0 a | 6666.7 ± 47.2 a |
7 | 7.6 ± 0.7 b | 39.2 ± 1.9 b | 20.2 ± 0.9 b | 33.1 ± 1.6 b | 346.7 ± 6.4 b | |
10 | 4.8 ± 0.2 c | 18.4 ± 2.2 c | 26.3 ± 0.6 a | 50.5 ± 1.1 c | 208.0 ± 1.3 c | |
Napin | 3 | 24.1 ± 0.7 a | NA | NA | 26.3 ± 1.3 a | 1239.3 ± 19.3 a |
7 | 27.5 ± 1.1 b | NA | NA | 26.9 ± 1.4 a | 103.6 ± 3.9 b | |
10 | 27.2 ± 0.7 a,b | NA | NA | 25.4 ± 0.6 a | 150.4 ± 1.6 c |
pH | Denaturation Temperature, Tm (°C) | Enthalpy (J/g) | Onset of Peak (°C) | End of Peak (°C) |
---|---|---|---|---|
3 | No peak was observed from 30 °C to 130 °C for both the species | |||
7 | 83.2 ± 0.8 a | 1.1 ± 0.3 a | 65–70 | 95–100 |
10 | 84.8 ± 0.2 a | 0.9 ± 0 a | 65–70 | 95–100 |
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Perera, S.P.; McIntosh, T.C.; Wanasundara, J.P.D. Structural Properties of Cruciferin and Napin of Brassica napus (Canola) Show Distinct Responses to Changes in pH and Temperature. Plants 2016, 5, 36. https://doi.org/10.3390/plants5030036
Perera SP, McIntosh TC, Wanasundara JPD. Structural Properties of Cruciferin and Napin of Brassica napus (Canola) Show Distinct Responses to Changes in pH and Temperature. Plants. 2016; 5(3):36. https://doi.org/10.3390/plants5030036
Chicago/Turabian StylePerera, Suneru P., Tara C. McIntosh, and Janitha P. D. Wanasundara. 2016. "Structural Properties of Cruciferin and Napin of Brassica napus (Canola) Show Distinct Responses to Changes in pH and Temperature" Plants 5, no. 3: 36. https://doi.org/10.3390/plants5030036