N-乙酰神经氨酸是最常见的一种唾液酸,通常位于细胞膜表面糖蛋白和糖脂的末端,具有多种生物学功能。由于其独特的生理和生化性质,N-乙酰神经氨酸在制药、化妆品和食品行业的应用越来越广泛,需求量不断增加。N-乙酰神经氨酸在大肠杆菌中的异源生物合成需要与中心碳代谢过程竞争前体物,因此,碳源利用效率,以及碳代谢流在细胞生长与N-乙酰神经氨酸合成中的分配,直接影响N-乙酰神经氨酸的合成效率。该研究从碳源利用调控角度,比较了葡萄糖、甘油,以及两者混合利用对N-乙酰神经氨酸合成的影响。在单一葡萄糖碳源条件下,菌株NEU5AC-2中pfkA的敲除减弱了中心碳代谢的强度,减少了副产物乙酸的积累,协调了前体物N-乙酰甘露糖胺与磷酸烯醇式丙酮酸的供应,5 L发酵罐分批补料发酵38 h产生23.8 g/L N-乙酰神经氨酸,获得了较高的发酵生产水平,具有较强的工业应用潜力。该研究可以为其他以糖酵解途径的中间代谢物为前体物的异源产物合成提供借鉴。
伊进行
,
宗媛
,
孙超
,
张晓雨
,
耿自豪
,
孙文超
,
张春月
,
刘政凯
,
熊雯雯
,
唐宇琳
,
马倩
. 大肠杆菌利用不同碳源生产N-乙酰神经氨酸的研究[J]. 食品与发酵工业, 2025
, 51(1)
: 18
-25
.
DOI: 10.13995/j.cnki.11-1802/ts.038677
N-acetylneuramic acid, the most common type of sialic acids, generally locates at the terminal of glycoprotein and glycolipid on the surface of cell membrane, where it performs a variety of biological functions.Because of its unique physiological and biochemical properties, it is more and more widely used in pharmaceutical, cosmetic and food industries, and its demand will continue to increase.The heterologous biosynthesis of N-acetylneuraminic acid in Escherichia coli needs to compete precursors with the process of central carbon metabolism.Therefore, the efficiency of carbon source utilization and the distribution of carbon metabolic flux in cell growth and N-acetylneuraminic acid synthesis directly affect the synthesis efficiency of N-acetylneuraminic acid.In this study, the effects of glucose, glycerol and their mixture on the synthesis of N-acetylneuraminic acid were compared from the point of view of carbon source utilization regulation.When using single glucose carbon source, the knockout of pfkA in strain NEU5AC-2 weakened the intensity of central carbon metabolism, reduced the accumulation of by-product acetic acid, and coordinated the supply of precursors GlcNAc and PEP.NEU5AC-2 produced 23.8 g/L N-acetylneuraminic acid after 38 h of fed-batch fermentation in a 5-L bioreactor, which obtained higher level of fermentation production and strong industrial application potential.This study shed light on similar studies using metabolites in EMP as precursors for the biosynthesis of heterologous chemicals.
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