文献类型：期刊文献

英文题名：De novo biosynthesis and nicotinamide biotransformation of nicotinamide mononucleotide by engineered yeast cells

作者：Ren, Yanna[1];Han, Bei[1];Wang, Shijie[1];Wang, Xingbin[1];Liu, Qi[1];Cai, Menghao[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai, Peoples R China;[3]Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, Shanghai, Peoples R China

年份：2024

卷号：17

期号：11

外文期刊名：MICROBIAL BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001368706400001)】；

基金：This study was supported by the National Natural Science Foundation of China (32201206), China Postdoctoral Science Foundation (2022M711146) and the National Key R&D Program of China (2023YFA0914102).

语种：英文

摘要：beta-Nicotinamide mononucleotide (NMN) is a precursor of NAD+ in mammals. Research on NAD+ has demonstrated its crucial role against aging and disease. Here two technical paths were established for the efficient synthesis of NMN in the yeast Pichia pastoris, enabling the production of NMN from the low-cost nicotinamide (NAM) or basic carbon sources. The yeast host was systematically modified to adapt to the biosynthesis and accumulation of NMN. To improve the semi-biosynthesis of NMN from NAM, nicotinamide phosphoribosyltransferases were expressed intracellular to evaluate their catalytic activities. The accumulation of extracellular NMN was further increased by the co-expression of an NMN transporter. Fine-tuning of gene expression level produced 72.1 mg/L NMN from NAM in flasks. To achieve de novo biosynthesis NMN, a heterologous biosynthetic pathway was reassembled in yeast cells. Fine-tuning of pathway nodes by the modification of gene expression level and enhancement of precursor generation allowed efficient NMN synthesis from glucose (36.9 mg/L) or ethanol (57.8 mg/L) in flask. Lastly, cultivations in a bioreactor in fed-batch mode achieved an NMN titre of 1004.6 mg/L at 165 h from 2 g NAM and 868 g glucose and 980.4 mg/L at 91 h from 160 g glucose and 557 g ethanol respectively. This study provides a foundation for future optimization of NMN biosynthesis by engineered yeast cell factories.