Production of 2′-fucosyllactose using ⍺1,2-fucosyltransferase from a GRAS bacterial strain

Abstract

2′-Fucosyllactose (2′-FL) is a major oligosaccharide found in human breast milk. It is produced from GDP-L-fucose and D-lactose by ⍺1,2-fucosyltransferase (⍺1,2-fucT), but the enzyme has been identified mostly in pathogens. In this study, an ⍺1,2-fucT was isolated from a Generally Recognized as Safe (GRAS) Bacillus megaterium strain. The enzyme was successfully expressed in metabolically-engineered Escherichia coli. Furthermore, replacement of non-conserved amino acid residues with conserved ones in the protein led to an increase in the rate of 2′-FL production. As a result, fed-batch fermentation of E. coli produced 30 g/L of 2′-FL from glucose and lactose. Thus, the overproduction of 2′-FL using a novel enzyme from a GRAS bacteria strain was successfully demonstrated.