Issue 28, 2025

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

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, and other critical metabolic pathways. An NADH synthesis pathway in artificial cells starting from D-ribose was constructed with a 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 the NADH synthesis pathway lays the foundation for fabricating artificial cells with complicated metabolic networks.

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

Supplementary files

Article information

Article type
Edge Article
Submitted
01 Feb 2025
Accepted
05 Jun 2025
First published
06 Jun 2025
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2025,16, 12969-12981

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

Y. Liu, S. Du, X. Zhang, C. Li, S. Li, W. Xu, J. Zhao, W. Mu and X. Han, Chem. Sci., 2025, 16, 12969 DOI: 10.1039/D5SC00852B

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements