A combined pseudouridine biomanufacturing platform enabled by a streamlined designer pathway.

IF 15.7 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Nature Communications Pub Date : 2025-10-06 DOI:10.1038/s41467-025-63906-0

Le Yu,Ruyi Chen,Chenyue Zhang,Zhengyuan Wang,Zhuqing Wang,Xinyue Zeng,Han Liang,Yuanyuan He,Yixuan She,Yifei Wang,Rong Gong,Xuemin Song,Zixin Deng,Qiang Fei,Wenqing Chen

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引用次数: 0

Abstract

mRNA vaccines, featured by incorporated pseudouridine (Ψ), represent a milestone in combating diseases, thus highlighting Ψ importance in drug development. However, economic and environmental challenges have persisted in sustainable Ψ production. Here, we formulate a streamlined designer Ψ pathway, comprising UMP nucleosidase, ΨMP glycosidase, and ΨMP phosphatase, and realize its gram-scale production by targeted discovery of a prominent UMP-preferred nucleosidase (NmYgdH). The optimized pathway, containing NmYgdH, RjPsuG (ΨMP glycosidase), and HDHD1 (ΨMP-specific phosphatase) is cloned into E. coli and systematic evaluation of multiple strategies achieves a Ψ titer of 44.8 g·L-1. Moreover, a thyA-dependent, tunable, and eco-friendly strategy for sustainable Ψ production is demonstrated in a 5 L bioreactor achieving titer of 45.3 g·L-1. Finally, we establish a simplified-strategy for rapid Ψ purification with a recovery-rate of 71%, and techno-economic analysis is employed to validate the feasibility and advantages of this fermentation platform for Ψ biomanufacturing. Therefore, this study provides a blueprint for industrial-production of nucleoside-related molecules.

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一个由流线型设计途径实现的组合假尿嘧啶生物制造平台。

以掺入假尿嘧啶（Ψ）为特征的mRNA疫苗是防治疾病的一个里程碑，因此突出了Ψ在药物开发中的重要性。然而，经济和环境挑战在可持续Ψ生产中持续存在。在这里，我们制定了一个流线型设计Ψ途径，包括UMP核苷酶，ΨMP糖苷酶和ΨMP磷酸酶，并通过有针对性地发现一个突出的UMP首选核苷酶（NmYgdH）来实现其克级生产。优化后的途径含有NmYgdH、RjPsuG （ΨMP糖苷酶）和HDHD1 （ΨMP-specific磷酸酶），将其克隆到大肠杆菌中，多种策略的系统评价达到Ψ滴度44.8 g·L-1。此外，一个依赖于thya的、可调的、环保的可持续Ψ生产策略在一个5l的生物反应器中得到了验证，滴度达到45.3 g·L-1。最后，我们建立了一个简化的Ψ快速纯化策略，回收率为71%，并通过技术经济分析验证了该发酵平台用于Ψ生物制造的可行性和优势。因此，本研究为核苷相关分子的工业化生产提供了蓝图。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nature Communications Biological Science Disciplines-

CiteScore

24.90

自引率

2.40%

发文量

6928

审稿时长

3.7 months

期刊介绍： Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.