التفاصيل البيبلوغرافية
| العنوان: |
Multi-strategy orthogonal enhancement and analysis of aldo-keto reductase thermal stability. |
| المؤلفون: |
Zhang, Lingzhi1 (AUTHOR), Zhou, Rui2 (AUTHOR), Liu, Dekai1 (AUTHOR), Zhu, Meinan1 (AUTHOR), Zhang, Guangya1 (AUTHOR), Zhang, Lijuan3 (AUTHOR), Zhou, Shu-Feng1 (AUTHOR) szhou@hqu.edu.cn, Jiang, Wei1 (AUTHOR) wjiang@hqu.edu.cn |
| المصدر: |
International Journal of Biological Macromolecules. Apr2024:Part 2, Vol. 264, pN.PAG-N.PAG. 1p. |
| مصطلحات موضوعية: |
*THERMAL stability, *PROTEIN engineering, *PROTEIN folding, *DRUG synthesis, *INTEGRAL domains |
| مستخلص: |
Given their outstanding efficiency and selectivity, enzymes are integral in various domains such as drug synthesis, the food industry, and environmental management. However, the inherent instability of natural enzymes limits their widespread industrial application. In this study, we underscore the efficacy of enhancing protein thermal stability through comprehensive protein design strategies, encompassing elements such as the free energy of protein folding, internal forces within proteins, and the overall structural design. We also demonstrate the efficiency and precision of combinatorial screening in the thermal stability design of aldo-keto reductase (AKR7-2-1). In our research, three single-point mutations and five combinatorial mutations were strategically introduced into AKR7-2-1, using multiple computational techniques. Notably, the E12I/S235I mutant showed a significant increase of 25.4 °C in its melting temperature (Tm). Furthermore, the optimal mutant, E12V/S235I, maintained 80 % of its activity while realizing a 16.8 °C elevation in Tm. Remarkably, its half-life at 50 °C was increased to twenty times that of the wild type. Structural analysis indicates that this enhanced thermal stability primarily arises from reduced oscillation in the loop region and increased internal hydrogen bonding. The promising results achieved with AKR7-2-1 demonstrate that our strategy could serve as a valuable reference for enhancing the thermal stability of other industrial enzymes. • Thermal Stability Enhancement: Applied protein design strategies focused on folding energy and structural dynamics. • Significant Thermal Improvements: E12I/S235I and E12V/S235I mutations led to notable increases in Tm and retained activity. • Simulation Insights: Enhanced stability shown by molecular dynamics through better hydrogen bonding and loop rigidity. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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