食品与发酵工业2025，Vol.51Issue(22)：51-59,9.DOI:10.13995/j.cnki.11-1802/ts.041876
筛选高活性烟酰胺磷酸核糖基转移酶强化烟酰胺单核苷酸的合成
Screening of highly active nicotinamide phosphoribosyltransferase to enhance nicotinamide mononucleotide synthesis
摘要
Abstract
Nicotinamide mononucleotide(NMN)is a derivative of the vitamin B group,which naturally exists in fruits,vegetables,and meat.It has been widely used in the field of food,health care products,and degenerative diseases treatment.Nicotinamide phosphori-bosyltransferase(NAMPT)is a rate-limiting enzyme in the NMN salvage synthesis pathway.The best NAMPT in current research is Vp-NadV derived from Vibrio bacteriophage,which has several defects,such as poor activity and low substrate utilization.In addition,its crystal structure has not been fully resolved.Based on the NMN fluorescence reaction system,this study aimed to construct a high-through-put screening system,and then to screen highly active VpNadV mutants combined with enzyme engineering,finally to apply for efficient synthesis of NMN in Escherichia coli.Through the high-throughput screening and enzyme engineering,a mutant with high enzymatic activi-ty,VpNadVL139V+R382G,was selected,which enzyme activity was 1.46-fold than that of the wild type.Based on the mutant,3.68 g/L NMN was achieved in shake flasks,and 24.4 g/L NMN was obtained after scaling up in a 5 L fermenter.In this study,a high-throughput screening system for Nicotinamide phosphoribosyltransferase(NAMPT)mutants was constructed,and then the catalytic mechanism of Vp-NadV was briefly analyzed,and finally the positive effect of NMN synthesis by mutants was further verified by fermentation,providing ide-as for the efficient production of NMN and nicotinamide derivatives.关键词
NAMPT/NMN/酶工程改造/高通量筛选/生物合成Key words
nicotinamide phosphoribosyltransferase(NAMPT)/nicotinamide mononucleotide(NMN)/enzyme engineering/high-throughput screening/biosynthesis引用本文复制引用
李展颜,王科,曾伟主,周景文..筛选高活性烟酰胺磷酸核糖基转移酶强化烟酰胺单核苷酸的合成[J].食品与发酵工业,2025,51(22):51-59,9.基金项目
国家重点研发计划项目(2023YFA0914500) （2023YFA0914500）
