相关实验视频

Updated: Jun 21, 2026

High Throughput Screening of Fungal Endoglucanase Activity in Escherichia coli

High Throughput Screening of Fungal Endoglucanase Activity in Escherichia coli

Published on: August 13, 2011

通过可扩展的高通量选平台对α1,2-fucosyltransferase的定向演化

Linhui Peng¹, Wenwen Yu², Xianhao Xu²

¹Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi, 214122, China; Institute of Future Food Technology, JITRI, Yixing, 214200, China.

International journal of biological macromolecules

|October 20, 2025

概括

研究人员开发了一种全细胞生物传感器和液滴微流体平台,用于选具有增强2'-糖乳糖 (2'-FL) 生产的α1,2-Fucosyltransferase (α1,2-FucT) 变体. 这种方法发现了一种高级突变物V93I,显著提高了催化效率.

科学领域:

生物技术是生物技术.
酵素工程是什么意思酵素工程
合成生物学合成生物学

背景情况:

α1,2-Fucosyltransferase (α1,2-FucT) 对于合成2'-富可酸乳糖 (2'-FL) 是至关重要的.
自然的α1,2-FucT表现出有限的催化活性,阻碍了它的工程.
缺乏有效的选平台来提高α1,2-FucT的性能.

研究的目的:

为工程 α1,2-FucT.开发一个高通量选平台.
确定具有显著增强2'-FL生物合成的催化活性的α1,2-FucT变体.
建立适用于其他甘氨基基转移酶的新型生物传感器战略.

主要方法:

设计了一个全细胞生物传感器,将2'-FL产量与光信号联系起来.
包含一个热敏的乳糖降解途径,以减轻乳糖干扰.
利用滴滴微流体技术对一个大型突变库 (0.1百万) 的高通量查.

主要成果:

成功开发了一个跨尺度滴滴微流体选平台.
确定了Helicobacter pylori α1,2-FucT的V93I突变,其催化效率 (kcat/Km) 增加了2.31倍.
与野生类型相比,V93I突变体表现出明显改善的2'-FL合成.

结论:

开发的全细胞生物传感器和液滴微流体平台可实现高效的高通量选糖转移酶.
这一战略成功地确定了一种工程化α1,2-FucT变体,具有增强的2'-FL生产能力.
该平台显示了在选和设计其他糖转移酶方面广泛应用的潜力.

关键词:

定向进化是指导进化的.滴滴分类滴滴分类高通量选的高通量选全细胞生物传感器 α1,2-fucosyl转移酶是什么

更多相关视频

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

相关概念视频

Pharmacogenetics of Phase II Enzymes: N-acetyltransferase, Thiopurine S-methyltransferase, UDP-glucuronosyltransferase

Pharmacogenetics of Phase II Enzymes: N-acetyltransferase, Thiopurine S-methyltransferase, UDP-glucuronosyltransferase

Phase II biotransformation reactions are essential for detoxifying and eliminating xenobiotics, including many pharmaceutical compounds. These reactions typically involve conjugation, the covalent attachment of polar endogenous groups such as glucuronic acid, sulfate, methyl, or acetyl moieties to functional groups introduced during Phase I metabolism. The resulting conjugates are more water-soluble, enabling efficient renal or biliary excretion.The major classes of Phase II enzymes include...