Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli
Abstract
:1. Introduction
2. Results
2.1. Construction of Random Mutant Library and Screening for Salt-Tolerance Variants
2.2. EP-5 Significantly Improved the Salt Tolerance to E. coli Transformant
2.3. EP-5 Conferred Other Abiotic Tolerances to E. coli Transformants
2.4. S32P Mutation Changed the Relationship of Two α-Helixes on N- and C-Terminal in EP-5
2.5. EP-5 Maintained the Normal Cell Morphology of E. coli Transformant
2.6. EP-5 Improved the Antioxidant Capacity of E. coli Transformant
2.7. EP-5 Stimulated NO Synthesis in E. coli Transformant
3. Discussion
4. Materials and Methods
4.1. Plasmid, Chemical Reagents and Strains
4.2. Construction of Ds-26-16 Random Mutation Library
4.3. Screening for Ds-26-16 Variant
4.4. Stress Tolerance Analysis
4.5. Homology Modeling
4.6. Cell Surface Scanning with SEM
4.7. Antioxidant Enzyme Activity, MDA and NO Content Analysis
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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NaCl (M) | pET-21b(+) | Ds-16-16 | EP-5 |
---|---|---|---|
0 | 2.23 ± 0.61 a | 2.37 ± 0.74 a | 2.32 ± 0.86 a |
0.51 | 31.25 ± 9.84 a | 2.70 ± 0.87 b | 2.36 ± 0.72 b |
1.03 | 7.29 ± 6.83 a | 4.20 ± 1.72 b | 4.77 ± 2.95 b |
1.54 | 1.81 ± 1.22 c | 3.06 ± 1.78 b | 3.74 ± 2.31 a |
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Guo, Y.; Dong, Y.; Hong, X.; Pang, X.; Chen, D.; Chen, X. Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli. Int. J. Mol. Sci. 2016, 17, 1813. https://doi.org/10.3390/ijms17111813
Guo Y, Dong Y, Hong X, Pang X, Chen D, Chen X. Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli. International Journal of Molecular Sciences. 2016; 17(11):1813. https://doi.org/10.3390/ijms17111813
Chicago/Turabian StyleGuo, Yuan, Yanping Dong, Xiao Hong, Xiaonan Pang, Defu Chen, and Xiwen Chen. 2016. "Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli" International Journal of Molecular Sciences 17, no. 11: 1813. https://doi.org/10.3390/ijms17111813