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Multi-strategy orthogonal enhancement and analysis of aldo-keto reductase thermal stability.
Zhang, Lingzhi; Zhou, Rui; Liu, Dekai; Zhu, Meinan; Zhang, Guangya; Zhang, Lijuan; Zhou, Shu-Feng; Jiang, Wei.
Afiliación
  • Zhang L; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China.
  • Zhou R; Shanghai Marine Diesel Engine Research Institute, Shanghai 201108, PR China.
  • Liu D; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China.
  • Zhu M; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China.
  • Zhang G; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China.
  • Zhang L; State Key Laboratory of Microbial Technology, Shandong University, No. 72 Binhai Road, Qingdao 266237, PR China.
  • Zhou SF; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China. Electronic address: szhou@hqu.edu.cn.
  • Jiang W; College of Chemical Engineering, Huaqiao University, Xiamen 361021, Fujian Province, PR China. Electronic address: wjiang@hqu.edu.cn.
Int J Biol Macromol ; 264(Pt 2): 130691, 2024 Apr.
Article en En | MEDLINE | ID: mdl-38458293
ABSTRACT
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mutación Puntual Idioma: En Revista: Int J Biol Macromol Año: 2024 Tipo del documento: Article

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Mutación Puntual Idioma: En Revista: Int J Biol Macromol Año: 2024 Tipo del documento: Article
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