在UDP-Galactose 4-Epimerase中的工程酸性抵抗使得从糖中有效的UDP-Galactose合成成为可能
Xinyu Liu1,2, Zhongbao Ma1,2, Yu Shen1
1School of Biotechnology and Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China.
Journal of agricultural and food chemistry
|March 26, 2025
概括
设计了一种耐酸酶变体 (GALEM2) 来克服UDP-Gal合成中的pH冲突. 这一突破使得使用级联催化从糖中快速,高效地生产尿素二酸银.
科学领域:
- 生物催化剂是一种生物催化剂.
- 酶工程是什么? 酶工程是什么?
- 碳水化合物生物合成
背景情况:
- 尿素二酸银糖 (UDP-Gal) 对于碳水化合物生物合成至关重要,但受供应和成本的限制.
- 现有用于UDP-Gal合成的两种酶级联系统面临着UDP-银4-表皮酶 (GALE) 和糖糖合成酶 (Susy) 之间的pH冲突,阻碍了生产.
研究的目的:
- 设计GALE的表面电荷变体,增强抗酸性,以解决UDP-Gal合成中的pH冲突.
- 通过级联催化提高UDP-Gal生产的效率和减少反应时间.
主要方法:
- 使用表面电荷工程来创建一种抗酸的GALE变体 (GALEM2).
- 在两种酶级联系统中使用了工程GALEM2和修改的糖合成酶 (SusyM6).
- GALEM2的表征包括在酸性条件下评估酶活性,半衰期和pH电阻.
主要成果:
- GALEM2表现出显著改善的酶活性 (214.26%的野生类型在pH6.5) 和延长的半衰期 (增加2小时在pH6.5).
- 由于局部表面电荷重塑和降低同电点,工程酶表现出增强的pH电阻.
- 使用GALEM2和SusyM6的级联系统在1.25小时内实现了UDP-Gal度24.5mM,时空收益率为12g/L/h.
结论:
- 表面充电工程有效地提高了GALE在酸性条件下的抗酸性和催化性能.
- 开发的级联催化路径提供了一种强大,高效和快速的方法,用于从糖糖中合成UDP-Gal.
- 这种工程酶系统有望在碳水化合物生物合成中大规模应用UDP-Gal.
相关概念视频
Oligosaccharide Assembly
2.7K
Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
Multiple sugar molecules that may or may...
2.7K
Biosynthesis of Polysaccharides
990
Polysaccharides such as glycogen and starch are synthesized from nucleoside diphosphate sugars, primarily uridine diphosphate glucose (UDPG) and adenosine diphosphate glucose (ADPG). These activated glucose donors act as key intermediates in carbohydrate metabolism and biosynthesis. UDPG primarily involves glycogen synthesis in animals and many bacteria, while ADPG plays a fundamental role in starch synthesis in plants and certain bacteria.UDPG is formed when glucose-1-phosphate reacts with...
990
Peptidoglycan Synthesis
4.8K
Structure of PeptidoglycanPeptidoglycan is a vital structural component of the bacterial cell wall, providing mechanical strength and shape to the cell. It consists of repeating units of two sugars—N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM)—linked by β-1,4 glycosidic bonds. These sugar chains are cross-linked by short peptide chains, forming a mesh-like polymer that surrounds the bacterial plasma membrane.Cytoplasmic Phase – Precursor SynthesisPeptidoglycan...
4.8K
Inducible Operons: lac Operon
3.1K
The lac operon in Escherichia coli is a model for understanding inducible gene regulation and metabolic flexibility. It integrates local control by lactose and global regulation through catabolite repression, enabling E. coli to preferentially metabolize glucose when available and switch to lactose utilization when glucose is scarce.Structure and Function of the lac OperonThe lac operon contains three structural genes: lacZ (β-galactosidase), lacY (lactose permease), and lacA...
3.1K


