Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis of MIONPs and APTES-Coated MIONPs
2.2. Immobilization of Recombinant CrCLH1
2.3. Immobilization Yield and Enzyme Capacity
2.4. Effects of Optimal pH and pH Stability of the Free and Immobilized Enzymes
2.5. Optimal Temperature and Thermostability of Free and Immobilized Enzymes
2.6. Operational Stability
3. Materials and Methods
3.1. Materials
3.2. Synthesis of MIONPs
3.3. APTES-Coated MIONPs
3.4. Characterization of MIONPs and APTES-Coated MIONPs
3.5. Synthesis of Recombinant CrCLH1-APTES-Coated MIONPs
3.6. Preparation of Enzymes
3.7. Chlase Assay of the Immobilized Enzyme
3.8. Biochemical Analyses of the Free and Immobilized Enzymes
3.9. Operational Stability
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Samples | Amount of Protein Binding (mg/g Gel) | Immobilization Yield (% b) | Enzymatic Activity (U c/g Gel) | Specific Activity of Immobilized Enzymes (U/mg Protein) | Specific Activity of Non-Immobilized Enzymes (U/mg Protein) |
---|---|---|---|---|---|
Free CrCLH1 | 0 | ND a | ND | ND | 186 ± 22 |
Immobilized CrCLH1 | 32.1 ± 0.3 | 98.99 ± 0.91 | 722.3 ± 50.3 | 131.0 ± 9.1 | 4.1 ± 0.3 |
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Yen, C.-C.; Chuang, Y.-C.; Ko, C.-Y.; Chen, L.-F.O.; Chen, S.-S.; Lin, C.-J.; Chou, Y.-L.; Shaw, J.-F. Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives. Molecules 2016, 21, 972. https://doi.org/10.3390/molecules21080972
Yen C-C, Chuang Y-C, Ko C-Y, Chen L-FO, Chen S-S, Lin C-J, Chou Y-L, Shaw J-F. Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives. Molecules. 2016; 21(8):972. https://doi.org/10.3390/molecules21080972
Chicago/Turabian StyleYen, Chih-Chung, Yao-Chen Chuang, Chia-Yun Ko, Long-Fang O. Chen, Sheau-Shyang Chen, Chia-Jung Lin, Yi-Li Chou, and Jei-Fu Shaw. 2016. "Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives" Molecules 21, no. 8: 972. https://doi.org/10.3390/molecules21080972