加入硫胺素的动态调控促进脂质体耶氏菌1,4-丁二醇的新合成。

IF 3.7 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
ACS Synthetic Biology Pub Date : 2025-07-18 Epub Date: 2025-07-03 DOI:10.1021/acssynbio.5c00015
Hongwei Guo, Yaqing Zeng, Mengqi Zhu, Tianqiu Huang, Weigao Wang, Catherine Madzak, Jun Zhao, Xiaohui Sun, Hongwen Chen, Guo Chen
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引用次数: 0

摘要

1,4-丁二醇(1,4- bdo)是最重要的平台二醇之一,作为共聚单体被广泛应用,每年生产数百万吨聚合物。微生物合成非天然的1,4- bdo是极具挑战性的。虽然在大肠杆菌中已经提出并应用了几种人工途径,但其中α-酮戊二酸酯(α-KG)向人工途径的分支代谢通量被认为是最具热力学效率的解决方案。在天然α-KG和琥珀酸盐高产菌多脂耶氏菌中建立1,4- bdo合成路线具有很高的潜力。将辅酶A依赖性的1,4- bdo合成途径引入到脂质体Y. lipolytica中,并对其限速步骤进行了研究,优化了硫胺素的用量,提高了前体和辅因子的利用率,并删除了竞争步骤。结果表明,1,4- bdo的合成受gaba -琥珀酸分流的代谢通量和NADPH有效性的影响。此外,在细胞生长和1,4- bdo合成之间存在权衡。采用将遗传动态调控纳入正代谢修饰的组合策略,分批发酵模式下获得了6217.1 mg·L-1 1,4- bdo。该研究还证明了酵母细胞中TCA中间体生物合成非天然化学物质的高潜力和能力,并为重新平衡异源途径和中心代谢之间的代谢通量提供了新的线索。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Enhanced 1,4-Butanediol de Novo Synthesis in Yarrowia lipolytica by Incorporating Dynamic Regulation of Thiamine.

1,4-Butanediol (1,4-BDO), one of the most important platform diols, has been widely utilized as a comonomer to manufacture several million tons of polymers every year. Microbial synthesis of the non-natural 1,4-BDO is extremely challenging. Although several artificial routes have been proposed and applied in Escherichia coli, among them, the branching metabolic flux from α-ketoglutarate (α-KG) to the artificial route was considered the most thermodynamically efficient solution. Establishing a 1,4-BDO synthesis route in Yarrowia lipolytica, a native α-KG and succinate hyperproducer, should have high potential. A CoA-dependent 1,4-BDO synthesis route was introduced into Y. lipolytica, followed by the investigation of rate-limiting steps, optimization of the thiamine dosage, improvement of the precursor and cofactor availability, and deletion of the competition step. It was illustrated that 1,4-BDO synthesis was susceptible to metabolic throughput of the GABA-succinate shunt and NADPH availability. Also, there was a trade-off between cellular growth and 1,4-BDO synthesis. A combinational strategy, in which hereditary dynamic regulation of thiamine was incorporated into those positive metabolic modifications, was implemented, and 6217.1 mg·L-1 1,4-BDO was achieved under the batch fermentation model. This work also demonstrated the high potential and capacity for biosynthesis of non-nature chemicals from TCA intermediates in the yeast cells, and provided a novel clue to rebalance the metabolic flux between the heterologous pathway and central metabolism.

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来源期刊
CiteScore
8.00
自引率
10.60%
发文量
380
审稿时长
6-12 weeks
期刊介绍: The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.
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