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基于序列和结构的热稳定D-Allulose 3-Epimerase和计算机引导的蛋白质工程的采矿,以改善酶活性

Hongbin Qi¹, Tong Wang¹, Huimin Li¹

¹Key Laboratory of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, National Engineering Laboratory for Industrial Enzymes, Tianjin 300457, China.

Journal of agricultural and food chemistry

|November 16, 2023

概括

研究人员使用大数据挖掘发现了一种来自*Thermogutta terrifontis*的高度热稳定的D-allulose 3-epimerase (DAEase). 酶工程进一步增强了其用于工业应用的催化活性.

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?
生物催化剂是一种生物催化剂.

背景情况:

D-Allulose是一种功能性甜味剂,使用D-allulose 3-epimerase (DAEase) 来从果糖中合成.
现有的DAEases缺乏适应恶劣工业条件所需的稳定性,这限制了它们的实际使用.

研究的目的:

为工业应用发现一种具有增强热稳定的新型DAEase.
为了设计一种具有改善催化活性的DAEase变体.

主要方法:

以字系学为指导的大数据分析和自由能量计算,以确定热稳定的DAEases.
结构引导的虚拟选和组合性的活性场所和测试/代性和突变发生,用于酶工程.
引导分子动力学模拟来分析酶基质相互作用.

主要成果:

从*Thermogutta terrifontis*中确定了TtDAE作为358个候选物中最热稳定的,在70°C时半衰期为32小时.
改造的TtDAE变种M4在催化活性上增加了5.12倍.
分子动力学模拟揭示了M4中扩大的基质结合口袋,改善了酶-基质的适合性.

结论:

一种新的,高温稳定的DAEase被发现并为改进的工业应用而设计.
大数据挖掘和合理酶修饰的结合方法对于开发工业生物催化剂是有效的.

关键词:

D - 氨酸3 - 胺酶是什么?大数据采矿是什么意思在片选中半理性的设计.热稳定性热稳定性

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