Biochemical characterization of Helicobacter pylori α1-3-fucosyltransferase and its application in the synthesis of fucosylated human milk oligosaccharides

  • Carbohydr Res. 2019 Jul 1:480:1-6. doi: 10.1016/j.carres.2019.05.007.
Jing Bai  1 ,  Zhigang Wu  2 ,  Go Sugiarto  3 ,  Madhusudhan Reddy Gadi  4 ,  Hai Yu  3 ,  Yanhong Li  3 ,  Cong Xiao  4 ,  Alice Ngo  3 ,  Baohua Zhao  5 ,  Xi Chen  6 ,  Wanyi Guan  7
Affiliations
  • 1. College of Life Science, Hebei Normal University, Shijiazhuang, Hebei, 050024, China.
  • 2. College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang, Hebei, 050018, China.
  • 3. Department of Chemistry, University of California, One Shields Avenue, Davis, CA, 95616, USA.
  • 4. Department of Chemistry, Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA, 30303, USA.
  • 5. College of Life Science, Hebei Normal University, Shijiazhuang, Hebei, 050024, China. Electronic address: [email protected].
  • 6. Department of Chemistry, University of California, One Shields Avenue, Davis, CA, 95616, USA. Electronic address: [email protected].
  • 7. College of Life Science, Hebei Normal University, Shijiazhuang, Hebei, 050024, China. Electronic address: [email protected].
PMID: 31132553 DOI: 10.1016/j.carres.2019.05.007
Abstract

Fucosylated Human Milk Oligosaccharides (HMOs) have important biological functions. Enzymatic synthesis of such compounds requires robust fucosyltransferases. A C-terminal 66-amino acid truncated version of Helicobacter pylori α1-3-fucosyltransferase (Hp3FT) is a good candidate. Hp3FT was biochemically characterized to identify optimal conditions for enzymatic synthesis of fucosides. While N-acetyllactosamine (LacNAc) and lactose were both suitable acceptors, the former is preferred. At a low Guanosine 5'-diphospho-β-L-fucose (GDP-Fuc) to acceptor ratio, Hp3FT selectively fucosylated LacNAc. Based on these enzymatic characteristics, diverse fucosylated HMOs, including 3-fucosyllactose (3-FL), lacto-N-fucopentaose (LNFP) III, lacto-N-neofucopentaose (LNnFP) V, lacto-N-neodifucohexaose (LNnDFH) II, difuco- and trifuco-para-lacto-N-neohexaose (DF-paraLNnH and TF-para-LNnH), were synthesized enzymatically by varying the ratio of the donor and acceptor as well as controlling the order of multiple glycosyltransferase-catalyzed reactions.

Keywords
Enzymatic synthesis; Fucosyltransferase; Human milk oligosaccharide; Substrate specificity.
Products