基于能量和进化设计的inulosucrase用于增强热稳定性和胰岛素的生产
Thanapon Charoenwongpaiboon1, Karan Wangpaiboon2, Manatsanan Puangpathanachai2
1Department of Chemistry, Faculty of Science, Silpakorn University, Nakhon Pathom, 73000, Thailand. charoenwongpaib_t@su.ac.th.
Applied microbiology and biotechnology
|September 9, 2023
概括
工程化糖酶 (LrInu) 变种显示了增强的热稳定性和活性. 该G237P变种显著提高了胰岛素合成效率,使其成为工业应用的前景.
科学领域:
- 酶学 是一种酶学.
- 蛋白质工程是指蛋白质工程.
- 生物技术是生物技术.
背景情况:
- 来自Lactobacillus reuteri 121 (LrInu) 的胰岛素糖酶对于生产诸如胰岛素和果糖类之类的益生菌有价值.
- 由于LrInu的温度稳定性不足,其工业应用受到限制.
研究的目的:
- 通过计算设计和表征LrInu.的热稳定变体.
- 为了识别具有更好的稳定性和胰岛素合成能力的LrInu变体.
主要方法:
- 使用FireProt服务器对热稳定变体的计算预测.
- 理性选择,蛋白质表达和9个LrInu变体的表征.
- 谱学和分子动力学 (MD) 模拟来分析结构变化和稳定性.
主要成果:
- 与野生型LrInu.相比,G237P变种显示出显著增强的热稳定性和活性.
- G237P的最佳温度增加了10°C (达到60°C),半衰期延长了5倍以上.
- MD模拟表明结构刚度增加,本地接触有助于G237P的热稳定性.
- 与野生类型酶相比,G237P显示了更有效的胰岛素合成.
结论:
- 该G237P变体代表了一个非常有前途的热稳定性胰岛素糖酶,用于工业益生菌合成.
- 计算式蛋白质设计有效地改善了酶特性,例如热稳定性.
- 增强的LrInu变种可以导致更有效地生产有价值的益生菌化合物.
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