基于蛋白质表面电荷的计算机辅助合理设计策略，提高侏罗纪地杆菌新型酯酶的热稳定性。

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2024-06-01 Epub Date: 2024-03-25 DOI:10.1007/s10529-024-03473-4

Runfei Song, Jin Zhang, Mengyu Zhu, Lin Lin, Wei Wei, Dongzhi Wei

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引用次数: 0

摘要

目的：尽管革囊菌是重要的嗜热菌源，但目前还没有关于侏罗纪革囊菌中恒温酯酶基因的报道，也没有关于提高酯酶热稳定性的合理设计策略的报道：结果：基因 gju768 与 Geobacillus sp.利用吉布斯展开自由能（ΔΔG）计算器和突变位点到催化位点的距离（DsCα-Cα）来筛选合适的消除表面负电荷的突变位点，突变体（D24N、E221Q和E253Q）显示出稳定的突变体，其热稳定性高于野生型（WT）。突变体 E253Q 的热稳定性最好，65 ℃ 时的半衰期（T1/2）为 32.4 分钟，是 WT（17.9 分钟）的 1.8 倍：基因 gju768 的克隆和基于表面电荷工程的合理设计有助于从地杆菌中鉴定出恒温酯酶，并探索热稳定性的进化策略。

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Computer-aided rational design strategy based on protein surface charge to improve the thermal stability of a novel esterase from Geobacillus jurassicus.

Objectives: Although Geobacillus are significant thermophilic bacteria source, there are no reports of thermostable esterase gene in Geobacillus jurassicus or rational design strategies to increase the thermal stability of esterases.

Results: Gene gju768 showed a highest similarity of 15.20% to esterases from Geobacillus sp. with detail enzymatic properties. Using a combination of Gibbs Unfolding Free Energy (∆∆G) calculator and the distance from the mutation site to the catalytic site (Ds_Cα-Cα) to screen suitable mutation sites with elimination of negative surface charge, the mutants (D24N, E221Q, and E253Q) displayed stable mutants with higher thermal stability than the wild-type (WT). Mutant E253Q exhibited the best thermal stability, with a half-life (T_1/2) at 65 °C of 32.4 min, which was 1.8-fold of the WT (17.9 min).

Conclusion: Cloning of gene gju768 and rational design based on surface charge engineering contributed to the identification of thermostable esterase from Geobacillus sp. and the exploration of evolutionary strategies for thermal stability.

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.