Counteraction of stability-activity trade-off of Nattokinase through flexible region shifting

IF 9.8 1区农林科学 Q1 CHEMISTRY, APPLIED

Food Chemistry Pub Date : 2023-05-03 DOI:10.1016/j.foodchem.2023.136241

Jie Luo, Chenshuo Song, Wenjing Cui, Laichuang Han, Zhemin Zhou

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引用次数: 5

Abstract

The widespread trade-off between stability and activity severely limits enzyme evolution. Although some progresses have been made to overcome this limitation, the counteraction mechanism for enzyme stability-activity trade-off remains obscure. Here, we clarified the counteraction mechanism of the Nattokinase stability-activity trade-off. A combinatorial mutant M4 was obtained by multi-strategy engineering, exhibiting a 20.7-fold improved half-life; meanwhile, the catalytic efficiency was doubled. Molecular dynamics simulation revealed that an obvious flexible region shifting in the structure of mutant M4 was occurred. The flexible region shifting which contributed to maintain the global structural flexibility, was considered to be the key factor for counteracting the stability-activity trade-off. Further analysis illustrated that the flexible region shifting was driven by region dynamical networks reshaping. This work provided deep insight into the counteraction mechanism of enzyme stability-activity trade-off, suggesting that flexible region shifting would be an effective strategy for enzyme evolution through computational protein engineering.

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柔性区域移位对纳豆激酶稳定性-活性权衡的抵消

稳定性和活性之间广泛存在的权衡严重限制了酶的进化。尽管在克服这一限制方面取得了一些进展，但酶稳定性-活性权衡的对抗机制仍然不清楚。在此，我们阐明了纳豆激酶稳定性-活性权衡的对抗机制。通过多策略工程获得了组合突变体M4，其半衰期提高了20.7倍;同时，催化效率提高了一倍。分子动力学模拟结果表明，突变体M4的结构发生了明显的柔性区移位。灵活的区域转移有助于保持全球结构的灵活性，被认为是抵消稳定-活动权衡的关键因素。进一步分析表明，区域动态网络重塑驱动了柔性区域移动。这项工作为酶稳定性-活性权衡的抵消机制提供了深入的见解，表明通过计算蛋白质工程，灵活的区域转移将是酶进化的有效策略。

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来源期刊

Food Chemistry 工程技术-食品科技

CiteScore

16.30

自引率

10.20%

发文量

3130

审稿时长

122 days

期刊介绍： Food Chemistry publishes original research papers dealing with the advancement of the chemistry and biochemistry of foods or the analytical methods/ approach used. All papers should focus on the novelty of the research carried out.