| Autor: |
Huang, Jinsha, Xie, Xiaoman, Zheng, Wanlin, Xu, Li, Yan, Jinyong, Wu, Ying, Yang, Min, Yan, Yunjun |
| Předmět: |
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| Zdroj: |
Biotechnology for Biofuels & Bioproducts; 2/24/2024, Vol. 17 Issue 1, p1-18, 18p |
| Abstrakt: |
Background: Biodiesel, an emerging sustainable and renewable clean energy, has garnered considerable attention as an alternative to fossil fuels. Although lipases are promising catalysts for biodiesel production, their efficiency in industrial-scale application still requires improvement. Results: In this study, a novel strategy for multi-site mutagenesis in the binding pocket was developed via FuncLib (for mutant enzyme design) and Rosetta Cartesian_ddg (for free energy calculation) to improve the reaction rate and yield of lipase-catalyzed biodiesel production. Thermomyces lanuginosus lipase (TLL) with high activity and thermostability was obtained using the Pichia pastoris expression system. The specific activities of the mutants M11 and M21 (each with 5 and 4 mutations) were 1.50- and 3.10-fold higher, respectively, than those of the wild-type (wt–TLL). Their corresponding melting temperature profiles increased by 10.53 and 6.01 °C, T 50 15 (the temperature at which the activity is reduced to 50% after 15 min incubation) increased from 60.88 to 68.46 °C and 66.30 °C, and the optimum temperatures shifted from 45 to 50 °C. After incubation in 60% methanol for 1 h, the mutants M11 and M21 retained more than 60% activity, and 45% higher activity than that of wt–TLL. Molecular dynamics simulations indicated that the increase in thermostability could be explained by reduced atomic fluctuation, and the improved catalytic properties were attributed to a reduced binding free energy and newly formed hydrophobic interaction. Yields of biodiesel production catalyzed by mutants M11 and M21 for 48 h at an elevated temperature (50 °C) were 94.03% and 98.56%, respectively, markedly higher than that of the wt–TLL (88.56%) at its optimal temperature (45 °C) by transesterification of soybean oil. Conclusions: An integrating strategy was first adopted to realize the co-evolution of catalytic efficiency and thermostability of lipase. Two promising mutants M11 and M21 with excellent properties exhibited great potential for practical applications for in biodiesel production. Highlights: Biodiesel is a sustainable, renewable and clean energy alternative to fossil fuels. Multi-point mutants were created by first integrating FuncLib and Cartesian_ddg. M21 achieved 93.37% FAME yield in 24 h, superior to 88.56% of wt–TLL in 48 h. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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