Improving Biosynthesis Efficiency of Nicotinamide Mononucleotide by ATP Recycling Engineering and Condition Optimization

作者：Cun‐Duo Tang, Taisong Shen, Xueyang Bai, Yao Wang, Jiashu Zhang, Yunchao Kan, Dandan Li, Lunguang Yao, Hongling Shi · 发表于：Journal of Agricultural and Food Chemistry · 年份：2025 · DOI：10.1021/acs.jafc.5c03017 · 被引用次数：6 · 研究领域：CRISPR and Genetic Engineering、Biochemical and Molecular Research、Sirtuins and Resveratrol in Medicine

Nicotinamide mononucleotide (NMN) is a very important bioactive nucleotide that is of great help to human health. However, its widespread application has been limited by its high production costs, especially the cost of the core substrates, coenzyme, and enzymes. In this study, the ADP/GDP-polyphosphate phosphotransferase Rh PPK2 originating from Rhodobacter sphaeroides was successfully expressed in Escherichia coli with high-level solubility, and the enzyme activity in the lysate supernatant reached 21.9 ± 0.65 U/mL. And then, the temperature profiles, pH profiles, and kinetic parameters of purified re Rh PPK2 were systematically characterized, which demonstrate its potential for application in enzymatic ATP regeneration systems. Furthermore, the introduction of re Rh PPK2 for ATP regeneration significantly enhanced NMN production efficiency, achieving a 2.3-fold increase compared to the conventional ATP supplementation method. Finally, the production efficiency of NMN was further improved by a single-factor experiment and L9(3 4 ) orthogonal design, and the yield was up to 14.6 ± 0.51 g/L, about 5.4 times of the initial yield. This research substantially reduced NMN production costs and established a robust foundation for industrial-scale NMN production.