福建农业科技2025，Vol.56Issue(3)：38-44,7.DOI:10.13651/j.cnki.fjnykj.2025.03.006
大肠杆菌中β-烟酰胺单核苷酸的生物合成与条件优化
Biosynthesis and Condition Optimization of β-nicotinamide Mononucleotides in Escherichia coli
摘要
Abstract
In order to improve the production of β-NMN by the multi-enzyme cascade reaction of Escherichia coli with nicotinamide and D-ribose as the substrates.Based on the engineering bacterium B20 constructed in the laboratory,the recombinant bacterium B26 was constructed by replacing the key enzyme nicotinamide phosphoribosyl transferase(NAMPT).In order to improve its performance,the pH of the reaction system,the addition mode of niacinamide,the concentration of Mg2+,the starting substrate of polyphosphokinase(PPK)and the addition strategy of inorganic polyphosphate were optimized by using the single factor experiments.The results showed that there was no significant difference in the yield between the two engineering strains B20 and B26,but the biomass of the engineering strain B26 expressing the key enzyme CpNAMPTY15S was 1.78 times that of B20,and the total production of NMN increased by 71.6%.The results of the single factor experiment determined that in a 20 mL catalytic system containing a final concentration of 15 mmol·L-1 Mg2+,5 mmol·L-1 AMP,and 25 mmol·L-1 inorganic phosphate,the nicotinamide with a final concentration of 25 mmol·L-1 was added in batches at 0 h and 6 h,respectively,while maintaining the pH of 7.0 as the optimal conditions for the NMN catalyzed reaction with ribose and nicotinamide as the direct substrates.Under these conditions,the multi-enzyme cascade reaction was carried out for 12 h.The final NMN production reached 9.09 g·L-1.By screening the dominant strains and optimizing the reaction conditions,the production of NMN biosynthesized by Escherichia coli was effectively improved,which provided a cost-effective reference method for the large-scale production of NMN.关键词
大肠杆菌/β-烟酰胺单核苷酸/烟酰胺/核糖Key words
Escherichia coli/β-nicotinamide mononucleotide/Nicotinamide/D-ribose分类
生物学引用本文复制引用
翁萍萍,包添祺,卢蓓丝,郑晓音,杨欣伟..大肠杆菌中β-烟酰胺单核苷酸的生物合成与条件优化[J].福建农业科技,2025,56(3):38-44,7.基金项目
福建省自然科学基金面上项目(2021J01170). （2021J01170）
