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

Enzyme Microb Technol. 2023 Jun:167:110232. doi: 10.1016/j.enzmictec.2023.110232. Epub 2023 Mar 28.

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.

Keywords: 2′-Fucosyllactose; Bacillus megaterium; GRAS; Human milk oligosaccharide (HMO); ⍺1,2-fucosyltransferase.

MeSH terms

  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Fucosyltransferases* / analysis
  • Fucosyltransferases* / genetics
  • Fucosyltransferases* / metabolism
  • Humans
  • Lactose / metabolism
  • Milk, Human / chemistry
  • Oligosaccharides / metabolism
  • Trisaccharides

Substances

  • 2'-fucosyllactose
  • Fucosyltransferases
  • Lactose
  • Trisaccharides
  • Oligosaccharides