Screening of high-NMN-producing natural strains and biosynthesis of NMN using Nampt
DOI:
https://doi.org/10.18686/fnc.v1i1.7Keywords:
nicotinamide mononucleotide; fermentation conditions optimization; semi-rational designAbstract
Nicotinamide mononucleotide (NMN) is an endogenous substance in humans with high safety and thermal stability, and its application in cosmetics, medical health, and functional foods has received widespread attention. However, the synthesis process of NMN has problems, such as high cost, time-consuming process, and low yield, which limits the large-scale industrial application of NMN to a certain extent. Nicotinamide phosphoribosyl transferase (Nampt) is a critical enzyme in the technical route of the biological synthesis of NMN, which can catalyze the synthesis of NMN using nicotinamide and phosphoribosyl pyrophosphate. The screening and expression of Nampt with excellent enzymatic properties and stability is the key to the synthesis of NMN via this method. At present, the main problems in the technical route of NMN production using Nampt are that the catalytic activity of Nampt is low and the sources of Nampt are limited. In this study, we isolated Enterobacter chengduensis 2021T4.7, a microorganism with a high NMN production, and optimized its fermentation condition. The yield of NMN was up to 67.66 μM. In addition, we synthesized Nampt and constructed related recombinant high-yield engineered bacteria. We semi-rationally designed a Nampt structure derived from mice and obtained mutant mNampt-V365L with NMN yield as high as 135.99 μM, which increased by 62% from that of the wild type. Here, we screened high-NMN-yield natural strains and obtained high-NMN-yield strains through the semi-rational design optimization of Nampt enzymes, which provided new chassis microorganisms and new ideas for the conversion rate of NMN.
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Copyright (c) 2023 Chao Li, Zouguo Long, Haichao Zhang, Yuhui Lin, Liqing Zhao
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