Engineering of α-1,3-fucosyltransferases for production of 3-fucosyllactose in Escherichia coli.

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  • Additional Information
    • Source:
      Publisher: Academic Press Country of Publication: Belgium NLM ID: 9815657 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-7184 (Electronic) Linking ISSN: 10967176 NLM ISO Abbreviation: Metab Eng Subsets: MEDLINE
    • Publication Information:
      Original Publication: Brugge, Belgium ; Orlando, FL : Academic Press, c1999-
    • Subject Terms:
      Bacterial Proteins*/genetics ; Bacterial Proteins*/metabolism ; Escherichia coli*/enzymology ; Escherichia coli*/genetics ; Fucosyltransferases*/biosynthesis ; Fucosyltransferases*/genetics ; Lactose*/analogs & derivatives ; Lactose*/biosynthesis ; Lactose*/genetics ; Metabolic Engineering*; Helicobacter pylori/*genetics; Helicobacter pylori/enzymology ; Humans ; Protein Engineering
    • Abstract:
      Fucosyllactoses (FLs), present in human breast milk, have been reported to benefit human health immensely. Especially, 3-fucosyllactose (3-FL) has numerous benefits associated with a healthy gut ecosystem. Metabolic engineering of microorganisms is thought to be currently the only option to provide an economically feasible route for large-scale production of 3-FL. However, engineering principles for α-1,3-fucosyltransferases (1,3-FTs) are not well-known, resulting in the lower productivity of 3-FL than that of 2'-fucosyllactose (2'-FL), although both 2'-FL and 3-FL follow a common pathway to produce GDP-L-fucose. The C-terminus of 1,3-FTs is composed of heptad repeats, responsible for dimerization of the enzymes, and a peripheral membrane anchoring region. It has long been thought that truncation of most heptad repeats, retaining just 1 or 2, helps the soluble expression of 1,3-FTs. However, whether the introduction of truncated version of 1,3-FTs enhances the production of 3-FL in a metabolically engineered strain, is yet to be tested. In this study, the effect of these structural components on the production of 3-FL in Escherichia coli was evaluated through systematic truncation and elongation of the C-terminal regions of three 1,3-FTs from Helicobacter pylori. Although these three 1,3-FTs contained heptad repeats and membrane-anchoring regions of varying lengths, they commonly exhibited an optimal performance when the number of heptad repeats was increased, and membrane-binding region was removed. The production of 3-FL could be increased 10-20-fold through this simple strategy.
      (Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: Escherichia coli; Fucosyllactose; Fucosyltransferase; Human milk oligosaccharides
    • Accession Number:
      0 (Bacterial Proteins)
      EC 2.4.1.- (Fucosyltransferases)
      EC 2.4.1.152 (galactoside 3-fucosyltransferase)
      J2B2A4N98G (Lactose)
    • Publication Date:
      Date Created: 20180606 Date Completed: 20190722 Latest Revision: 20200930
    • Publication Date:
      20221213
    • Accession Number:
      10.1016/j.ymben.2018.05.021
    • Accession Number:
      29870790