Reprogramming of sugar transport pathways in Escherichia coli using a permeabilized SecY protein-translocation channel
Reprogramming of sugar transport pathways in Escherichia coli using a permeabilized SecY protein-translocation channel
复制标题
使用透化 SecY 蛋白易位通道重编程大肠杆菌中的糖转运途径
DOI：
10.1002/bit.27306
复制
发表时间：
2020-02-20
影响因子：
3.8
通讯作者：
Fan, Li-Hai
中科院分区：
文献类型：
--
作者：
Guo, Qiang;Mei, Sen;Fan, Li-Hai
在糖代谢的初始步骤中，糖特异性转运蛋白在糖通过质膜进入细胞质的过程中起着决定性作用。细菌中的 SecY 复合物 (SecYEG) 形成负责蛋白质易位的膜通道。目前的工作表明，透化的 SecY 通道可用作大肠杆菌中的非特异性糖转运蛋白。删除插头结构域的 SecY 允许葡萄糖、果糖、甘露糖、木糖和阿拉伯糖通过，并且通过额外的孔环突变，促进乳糖转运，表明通过透化 SecY 的糖通过与糖立体特异性无关。工程大肠杆菌在多种单糖上表现出快速生长，并受益于消除运输饱和、改善糖耐受性、减少竞争性抑制以及防止碳分解代谢物抑制，这些通常是天然糖摄取系统遇到的。 SecY 通道在原核生物中广泛存在，因此其他细菌也可以被改造为利用该系统来摄取糖。因此，SecY 通道为未来开发用于生物技术应用的强大细胞工厂提供了独特的糖通道。
In the initial step of sugar metabolism, sugar-specific transporters play a decisive role in the passage of sugars through plasma membranes into cytoplasm. The SecY complex (SecYEG) in bacteria forms a membrane channel responsible for protein translocation. The present work shows that permeabilized SecY channels can be used as nonspecific sugar transporters in Escherichia coli. SecY with the plug domain deleted allowed the passage of glucose, fructose, mannose, xylose, and arabinose, and, with additional pore-ring mutations, facilitated lactose transport, indicating that sugar passage via permeabilized SecY was independent of sugar stereospecificity. The engineered E. coli showed rapid growth on a wide spectrum of monosaccharides and benefited from the elimination of transport saturation, improvement in sugar tolerance, reduction in competitive inhibition, and prevention of carbon catabolite repression, which are usually encountered with native sugar uptake systems. The SecY channel is widespread in prokaryotes, so other bacteria may also be engineered to utilize this system for sugar uptake. The SecY channel thus provides a unique sugar passageway for future development of robust cell factories for biotechnological applications.