Intracellular Construction of Organelle-like Compartments Facilitates Metabolic Flux in Escherichia coli,Journal of Agricultural and Food Chemistry

当前位置： X-MOL 学术 › J. Agric. Food Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Intracellular Construction of Organelle-like Compartments Facilitates Metabolic Flux in Escherichia coli
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-09-04 , DOI: 10.1021/acs.jafc.4c06895
Li Wan ₁ , Juntao Ke ₁ , Yingying Zhu ₁ , Wenli Zhang ₁ , Wanmeng Mu ₁

Affiliation

The formation of well-designed synthetic compartments or membraneless organelles for applications in synthetic biology and cellular engineering has aroused enormous interest. However, establishing stable and robust intracellular compartments in bacteria remains a challenge. Here, we use the structured DIX domains derived from Wnt signaling pathway components, more specifically, Dvl2 and Axin1, as building blocks to generate intracellular synthetic compartments in Escherichia coli. Moreover, the aggregation behaviors and physical properties of the DIX-based compartments can be tailored by genetically embedding a specific dimeric domain into the DIX domains. Then, a pair of interacting motifs, consisting of the aforementioned dimeric domain and its corresponding binding ligand, was incorporated to modify the client recruitment pattern of the synthetic compartments. As a proof of concept, the human milk oligosaccharide lacto-N-tetraose (LNT) biosynthesis pathway was selected as a model metabolic pathway. The fermentation results demonstrated that the co-compartmentalization of sequential pathway enzymes into intracellular compartments created by DIX domain, or by the DIX domain in conjunction with interacting motifs, prominently enhanced the metabolic flux and increased LNT production. These synthetic protein compartments may provide a feasible and effective tool to develop versatile organelle-like compartments in bacteria for applications in cellular engineering and synthetic biology.

中文翻译：

细胞内类似细胞器的结构促进大肠杆菌的代谢通量

用于合成生物学和细胞工程应用的精心设计的合成区室或无膜细胞器的形成引起了人们的极大兴趣。然而，在细菌中建立稳定且坚固的细胞内区室仍然是一个挑战。在这里，我们使用源自 Wnt 信号通路组件的结构化 DIX 结构域，更具体地说，Dvl2 和 Axin1，作为构建块在大肠杆菌中生成细胞内合成区室。此外，基于 DIX 的区室的聚集行为和物理特性可以通过将特定的二聚结构域基因嵌入 DIX 结构域来定制。然后，掺入由上述二聚结构域及其相应的结合配体组成的一对相互作用基序，以修改合成区室的客户招募模式。作为概念验证，选择人乳寡糖乳-N-四糖（LNT）生物合成途径作为模型代谢途径。发酵结果表明，由 DIX 结构域或由 DIX 结构域与相互作用基序结合产生的连续途径酶共区室化到细胞内区室中，显着增强了代谢通量并增加了 LNT 产量。这些合成蛋白质区室可以提供一种可行且有效的工具来开发细菌中的多功能类细胞器区室，用于细胞工程和合成生物学的应用。

更新日期：2024-09-04

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南