Construction of the reduced nicotinamide adenine dinucleotide salvage pathway in artificial cells and its application in amino acid synthesis

IF 7.6 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Chemical Science Pub Date : 2025-06-06 DOI:10.1039/d5sc00852b

Yiming Liu, Shanshan Du, Xiangxiang Zhang, Chao Li, Shubin Li, Wenxia Xu, Jingjing Zhao, Wei Mu, Xiaojun Han

{"title":"Construction of the reduced nicotinamide adenine dinucleotide salvage pathway in artificial cells and its application in amino acid synthesis","authors":"Yiming Liu, Shanshan Du, Xiangxiang Zhang, Chao Li, Shubin Li, Wenxia Xu, Jingjing Zhao, Wei Mu, Xiaojun Han","doi":"10.1039/d5sc00852b","DOIUrl":null,"url":null,"abstract":"Reduced nicotinamide adenine dinucleotide (NADH) salvage pathway reconstitution is a crucial step toward autonomous artificial cells. In living systems, D-ribose is a fundamental precursor intricately involved in the synthesis of nucleotides, nucleic acids, and critical metabolic pathways. An NADH synthesis pathway in artificial cells starting from D-ribose was constructed with five-enzyme cascade containing ribokinase, ribose-phosphate pyrophosphokinase, nicotinamide phosphoribosyltransferase, nicotinamide mononucleotide adenylyltransferase, and formate dehydrogenase (RK, RPPK, NAMPT, NMNAT, and FDH), which efficiently converted 10 mM D-ribose into 415 μM NADH within 80 minutes under optimized conditions. The produced NADH was further used to drive the amino acid metabolism, i.e., to convert NH4+ and α-ketoglutarate to glutamate by introducing additional glutamate dehydrogenase (GDH) inside artificial cells. The successful reconstitution of NADH synthesis pathway lays the foundation for fabricate artificial cells with complicated metabolic net works.","PeriodicalId":9909,"journal":{"name":"Chemical Science","volume":"39 1","pages":""},"PeriodicalIF":7.6000,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chemical Science","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1039/d5sc00852b","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}

引用次数: 0

Abstract

Reduced nicotinamide adenine dinucleotide (NADH) salvage pathway reconstitution is a crucial step toward autonomous artificial cells. In living systems, D-ribose is a fundamental precursor intricately involved in the synthesis of nucleotides, nucleic acids, and critical metabolic pathways. An NADH synthesis pathway in artificial cells starting from D-ribose was constructed with five-enzyme cascade containing ribokinase, ribose-phosphate pyrophosphokinase, nicotinamide phosphoribosyltransferase, nicotinamide mononucleotide adenylyltransferase, and formate dehydrogenase (RK, RPPK, NAMPT, NMNAT, and FDH), which efficiently converted 10 mM D-ribose into 415 μM NADH within 80 minutes under optimized conditions. The produced NADH was further used to drive the amino acid metabolism, i.e., to convert NH4+ and α-ketoglutarate to glutamate by introducing additional glutamate dehydrogenase (GDH) inside artificial cells. The successful reconstitution of NADH synthesis pathway lays the foundation for fabricate artificial cells with complicated metabolic net works.

查看原文本刊更多论文

人工细胞中还原性烟酰胺腺嘌呤二核苷酸挽救通路的构建及其在氨基酸合成中的应用

还原烟酰胺腺嘌呤二核苷酸（NADH）挽救途径的重建是实现自主人工细胞的关键一步。在生命系统中，d -核糖是一个基本的前体，复杂地参与了核苷酸、核酸的合成和关键的代谢途径。通过核糖激酶、核糖-磷酸焦磷酸激酶、烟酰胺磷酸核糖基转移酶、烟酰胺单核苷酸腺苷基转移酶和甲酸脱氢酶（RK、RPPK、NAMPT、NMNAT和FDH）五酶级联构建了以d -核糖为起点的人工细胞NADH合成途径，在优化条件下，在80分钟内将10 mM的d -核糖高效转化为415 μM的NADH。产生的NADH进一步用于驱动氨基酸代谢，即通过在人工细胞内引入额外的谷氨酸脱氢酶（GDH）将NH4+和α-酮戊二酸转化为谷氨酸。NADH合成途径的成功重构为合成具有复杂代谢网络的人造细胞奠定了基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Chemical Science CHEMISTRY, MULTIDISCIPLINARY-

CiteScore

14.40

自引率

4.80%

发文量

1352

审稿时长

2.1 months

期刊介绍： Chemical Science is a journal that encompasses various disciplines within the chemical sciences. Its scope includes publishing ground-breaking research with significant implications for its respective field, as well as appealing to a wider audience in related areas. To be considered for publication, articles must showcase innovative and original advances in their field of study and be presented in a manner that is understandable to scientists from diverse backgrounds. However, the journal generally does not publish highly specialized research.