De novo biosynthesis of 2′-fucosyllactose in a metabolically engineered Escherichia coli using a novel ɑ1, 2-fucosyltransferase from Azospirillum lipoferum. (April 2023)

• Record Type: Journal Article
• Authors: Chen, Yihan; Zhu, Yingying; Wang, Hao; Chen, Roulin; Liu, Yuanlin; Zhang, Wenli; Mu, Wanmeng
• Is Part Of: Bioresource technology. Volume 374(2023)
• Publication Date: 2023-04
• Subjects: Human milk oligosaccharides -- 2′-Fucosyllactose -- ɑ1, 2-Fucosyltransferase -- Metabolic pathway flux -- Engineered strain; Biomass -- Periodicals; Biomass energy -- Periodicals; Bioremediation -- Periodicals; Agricultural wastes -- Periodicals; Factory and trade waste -- Periodicals; Organic wastes -- Periodicals; Bioénergie -- Périodiques; Déchets agricoles -- Périodiques; Déchets industriels -- Périodiques; Déchets organiques -- Périodiques; Déchets (Combustible) -- Périodiques; 662.88
• DOI: 10.1016/j.biortech.2023.128818
• Languages: English
• ISSNs: 0960-8524

Abstract

Graphical abstract: Highlights: A novel ɑ 1, 2-fucosyltransferase was used for 2′-fucosyllactose production. The yield of 2′-fucosyllactose was enhanced by introducing SAMT into strains. Only 2′-fucosyllactose can be produced by SAMT -based strains. The maximum of 2′-fucosyllactose reached 112.56 g/L by fed-batch cultivation. Abstract: Human milk oligosaccharides are complex, indigestible oligosaccharides that provide ideal nutrition for infant development. Here, 2′-fucosyllactose was efficiently produced in Escherichia coli by using a biosynthetic pathway. For this, both lacZ and wcaJ (encoding β -galactosidase and UDP-glucose lipid carrier transferase, respectively) were deleted to enhance the 2′-fucosyllactose biosynthesis. To further enhance 2′-fucosyllactose production, SAMT from Azospirillum lipoferum was inserted into the chromosome of the engineered strain, and the native promoter was replaced with a strong constitutive promoter (PJ23119 ). The titer of 2′-fucosyllactose was increased to 8.03 g/L by introducing the regulators rcsA and rcsB into the recombinant strains. Compared to wbgL -based strains, only 2′-fucosyllactose was produced in SAMT -based strains without other by-products. Finally, the highest titer of 2′-fucosyllactose reached 112.56 g/L in a 5 L bioreactor by fed-batch cultivation, with a productivity of 1.10 g/L/h and a yield of 0.98 mol/mol lactose, indicating a strong potential in industrial production.