Biochemical characterization of Helicobacter pylori α1–3-fucosyltransferase and its application in the synthesis of fucosylated human milk oligosaccharides. (1st July 2019)

• Record Type: Journal Article
• Authors: Bai, Jing; Wu, Zhigang; Sugiarto, Go; Gadi, Madhusudhan Reddy; Yu, Hai; Li, Yanhong; Xiao, Cong; Ngo, Alice; Zhao, Baohua; Chen, Xi; Guan, Wanyi
• Is Part Of: Carbohydrate research. Volume 480(2019)
• Page Start: 1
• Page End: 6
• Publication Date: 2019-07-01
• Subjects: Fucosyltransferase -- Substrate specificity -- Human milk oligosaccharide -- Enzymatic synthesis; Carbohydrates -- Periodicals; Chemistry, Organic -- Periodicals; Biochemistry -- Periodicals; Carbohydrates -- Periodicals; Chimie organique -- Périodiques; Glucides -- Périodiques; Biochemistry; Carbohydrates; Chemistry, Organic; Periodicals; Electronic journals; 507.78
• DOI: 10.1016/j.carres.2019.05.007
• Languages: English
• ISSNs: 0008-6215

Abstract

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. Graphical abstract: Image 1 Highlights: Helicobacter pylori α1–3-fucosyltransferase (Hp3FT) is biochemically characterized. Hp3FT prefers LacNAc over Lac as an acceptor substrate. Hp3FT selectively fucosylates LacNAc at a low GDP-Fuc to acceptor ratio. Selective fucosylation of Lac can be achieved by controlling glycosylation sequence. Diverse fucosylated HMOs have been synthesized using Hp3FT.