Methods for Enhancing the Thermal Stability of Alginate Lyase Using Different Computer-Aided Design Strategies
學生姓名:
賴昱亘
指導教授:
方翠筠
學期:
115上
摘 要:
Alginate lyases can degrade alginate through β-elimination reactions, producing alginate oligosaccharides with potential biological activity. However, the insufficient thermal stability of some natural alginate lyases limits their application in enzymatic processes. This report summarizes and compares two studies that used computer-aided rational design to improve the thermal stability of alginate lyases. The first study screened for mutation sites using HotSpot Wizard 3.0, PoPMuSiC, 18 B-factor, and ΔΔG analysis, identifying the Q246V and K249V mutants. Their half-lives at 50 °C increased to 3.85 and 3.65 h, respectively, with improved specific activity. The second study focused on the cold-adapted AlyC3 and combined PROSS and PDBePISA analyses to identify, through single-point and combinatorial mutations, the quadruple mutant A35P L95F-N138Y-M162I, whose melting temperature was 17.05 °C higher than that of the wild-type, and whose half-life at 50 °C was 8.21 times that of the wild-type. Both studies demonstrate that computer software can assist in screening for potential beneficial mutation sites and that experimental and structural analyses can be used to validate their thermal stability metrics, catalytic performance, and molecular mechanisms of action, thereby enhancing the applications of alginate lyases.
賴昱亘.pdf