Glycosyltransferase from Bacteroides gallinaceum Is a Novel α-1,3-Fucosyltransferase that Can Be Used for 3-Fucosyllactose Production In Vivo by Metabolically Engineered Escherichia coli

Abstract

As one of the attractive fucosylated human milk oligosaccharides, the biological production of 3-fucosyllactose (3-FL) has received great attention, as it exhibits many excellent physiological functions for infants. In this work, a novel 3-FL-producing α-1,3-fucosyltransferase (α1,3-FucT) named FutM2 from Bacteroides gallinaceum was first selected from nine potential candidates in the NCBI database. Then, a highly 3-FL-producing engineered Escherichia coli strain was constructed by modular pathway enhancement including the GDL-l-fucose precursor supply by overexpressing manC, manB, gmd, and wcaG (CBGW), and the 3-FL synthesis pathway by introducing B. gallinaceum FutM2. Finally, a titer of 20.3 g L-1 and productivity of 0.40 g L-1 h-1 of 3-FL were achieved in the 3-L bioreactor by engineered E. coli (ΔlacZΔwcaJ) harboring pCDF-CBGW and pET-futM2. Our study provided a novel α1,3-FucT from B. gallinaceum that could be used for 3-FL production, presenting an efficient microbial cell factory platform to de novo synthesize 3-FL from glycerol.