通过切断和定向进化的工程糖分异构酶,以获得热稳定性和高效的异构糖生产
Hao Meng1, Chao Li2, Qian Shen3
1Key Laboratory of Biometallurgy of Ministry of Education, School of Minerals Processing and Bioengineering, Central South University, Changsha 410083, P. R. China.
Journal of agricultural and food chemistry
|November 4, 2025
概括
改造的糖分异构酶 (SIase) 酶显示出对异构糖生产的增强活性和稳定性. 这项研究提出了一种改进的SIase,其性能得到了改进,使得高产量的糖糖转化为异黄糖.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 碳水化合物化学 碳水化合物化学
背景情况:
- 一种功能性糖,即异糖,是用糖异酶 (SIase) 来从糖中制成的.
- 实践中的SIase应用受到酶不稳定性和低活性的限制.
- 潘托亚分散SIase (PdSIase) 是一个改进的目标.
研究的目的:
- 为了增强Pantoea dispersa SIase (PdSIase) 的活性和热稳定性.
- 开发一个高通量选平台,用于SIase的定向演变.
- 为了促进大规模的异盐生物合成.
主要方法:
- 在PdSIase中切割灵活区域以创建PdSIaseΔ32.
- 使用高通量选平台进行定向进化.
- 结构建模和分子动力学模拟以了解稳定性增强.
- 使用工程 SIase 的方法将糖糖生物转化为异二醇糖.
主要成果:
- PdSIaseΔ32的活性增加了2.29倍.
- 定向进化产生了PdSIaseΔ32-V300D/D330T,其半衰期在45°C时提高了1.7倍.
- 增强的热稳定性归因于新的链间键.
- 实现了98.9%的高转化率 (791.2g/L从800g/L的糖糖中转化为异二醇).
结论:
- 工程 SIase 变体显示显著改善了活动和热稳定性.
- 一个强大的选平台加速了SIase的发展.
- 优化的SIase促进了高效和高产的异盐生产.
- 这些发现支持大规模的工业应用异醇生物合成.
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