Improvement of Catalytic Activity and Thermostability of Alginate Lyase VxAly7B-CM via Rational Computational Design Strategies DOI Creative Commons
Xin Ma, Ke Zhu, Kaiyang Wang

et al.

Marine Drugs, Journal Year: 2025, Volume and Issue: 23(5), P. 198 - 198

Published: May 1, 2025

Alginate lyase degrades alginate through the β-elimination mechanism to produce oligosaccharides (AOS) with notable biochemical properties and diverse biological activities. However, its poor thermostability limits large-scale industrial production. In this study, we employed a rational computational design strategy combining computer-aided evolutionary coupling analysis ΔΔGfold evaluation enhance both catalytic activity of VxAly7B-CM. Among ten single-point mutants, E188N S204G mutants exhibited increases in Tm from 47.0 °C 48.9 50.2 °C, respectively, specific activities 3701.02 U/mg 2812.01 at 45 °C. Notably, combinatorial mutant E188N/S204G demonstrated ΔTm 5 an optimal reaction temperature up 50 where reached 3823.80 U/mg—a 31% increase. Moreover, half-life was 38.4 h, which is 7.0 times that wild-type enzyme. Protein structural molecular dynamics simulations suggested enhanced performance may be attributed optimized surface charge distribution, strengthened hydrophobic interactions, increased tertiary structure stability. Overall, our findings provided valuable insights into enzyme stabilization strategies supported production functional AOS.

Language: Английский

Improvement of Catalytic Activity and Thermostability of Alginate Lyase VxAly7B-CM via Rational Computational Design Strategies DOI Creative Commons
Xin Ma, Ke Zhu, Kaiyang Wang

et al.

Marine Drugs, Journal Year: 2025, Volume and Issue: 23(5), P. 198 - 198

Published: May 1, 2025

Alginate lyase degrades alginate through the β-elimination mechanism to produce oligosaccharides (AOS) with notable biochemical properties and diverse biological activities. However, its poor thermostability limits large-scale industrial production. In this study, we employed a rational computational design strategy combining computer-aided evolutionary coupling analysis ΔΔGfold evaluation enhance both catalytic activity of VxAly7B-CM. Among ten single-point mutants, E188N S204G mutants exhibited increases in Tm from 47.0 °C 48.9 50.2 °C, respectively, specific activities 3701.02 U/mg 2812.01 at 45 °C. Notably, combinatorial mutant E188N/S204G demonstrated ΔTm 5 an optimal reaction temperature up 50 where reached 3823.80 U/mg—a 31% increase. Moreover, half-life was 38.4 h, which is 7.0 times that wild-type enzyme. Protein structural molecular dynamics simulations suggested enhanced performance may be attributed optimized surface charge distribution, strengthened hydrophobic interactions, increased tertiary structure stability. Overall, our findings provided valuable insights into enzyme stabilization strategies supported production functional AOS.

Language: Английский

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