Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside
Abstract
:1. Introduction
2. Results and Discussion
2.1. Expression and Purification of OsUGT79
2.2. Kinetic Characteristics of OsUGT79
Compound | Activity (%) |
---|---|
Control | 100 |
EDTA (1 mM) | 98 |
EDTA (5 mM) | 95 |
CaCl2 | 108 |
CuSO4 | ND |
MgSO4 | 117 |
MnCl2 | 143 |
MnSO4 | 139 |
ZnSO4 | ND |
FeSO4 | 114 |
2.3. UDP-Glucose Recycling and DON Production
2.4. Preparative Production and Purification of D3G
3. Experimental Section
3.1. Materials
3.2. Cloning of OsUGT79 and AtSUS1
3.3. Protein Expression and Purification
3.4. Glycosylation Assays
3.5. DON and D3G Determination by HPLC-MS/MS
3.6. Preparative High-Performance Liquid Chromatography
3.7. D3G Purity Analysis by HPLC-UV
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Michlmayr, H.; Malachová, A.; Varga, E.; Kleinová, J.; Lemmens, M.; Newmister, S.; Rayment, I.; Berthiller, F.; Adam, G. Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside. Toxins 2015, 7, 2685-2700. https://doi.org/10.3390/toxins7072685
Michlmayr H, Malachová A, Varga E, Kleinová J, Lemmens M, Newmister S, Rayment I, Berthiller F, Adam G. Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside. Toxins. 2015; 7(7):2685-2700. https://doi.org/10.3390/toxins7072685
Chicago/Turabian StyleMichlmayr, Herbert, Alexandra Malachová, Elisabeth Varga, Jana Kleinová, Marc Lemmens, Sean Newmister, Ivan Rayment, Franz Berthiller, and Gerhard Adam. 2015. "Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside" Toxins 7, no. 7: 2685-2700. https://doi.org/10.3390/toxins7072685