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Facile enzymatic conversion of lactose into lacto-N-tetraose and lacto-N-neotetraose

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Abstract

Lacto-N-tetraose (Galβ1-3GlcNAcβ1-3Galβ1-4Glc, LNT) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc, LNnT) were enzymatically synthesized by consecutive additions of GlcNAc and Gal residues to lactose. Lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc) was prepared first by the transfer of GlcNAc from UDP-GlcNAc to lactose by β-1,3-N-acetylglucosaminyltransferase from bovine serum. The resulting lacto-N-triose II was converted into LNT and LNnT utilizing two kinds of β-D-galactosidase-mediated transglycosylations. Thus, β-D-galactosidase from Bacillus circulans ATCC31382 induced regioselective galactosyl transfer from o-nitrophenyl β-D-galactoside to the OH-3″ position of lacto-N-triose II, and commercially available β-D-galactosidase from B. circulans to the OH-4″ position of lacto-N-triose II. These convenient processes are suitable for large-scale preparations of LNT and LNnT. As another method, LNT was directly synthesized from lactose as an initial substance, utilizing lacto-N-biosidase (Aureobacterium sp. L-101)-mediated transglycosylation with Galβ1-3GlcNAcβ-pNP donor.

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Authors and Affiliations

  1. Department of Applied Biological Chemistry, Shizuoka University, 836, Ohya, Shizuoka, 422-8529, Japan

    Takeomi Murata, Takahiro Inukai, Masanori Suzuki & Taichi Usui

  2. Numazu Industrial Research Institute of Shizuoka Prefecture, 3981-1 Ohoka, Numazu, Shizuoka, 424-1221, Japan

    Masaaki Yamagishi

Authors
  1. Takeomi Murata
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  2. Takahiro Inukai
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  3. Masanori Suzuki
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  4. Masaaki Yamagishi
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  5. Taichi Usui
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Correspondence to Taichi Usui.

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Murata, T., Inukai, T., Suzuki, M. et al. Facile enzymatic conversion of lactose into lacto-N-tetraose and lacto-N-neotetraose. Glycoconj J 16, 189–195 (1999). https://doi.org/10.1023/A:1007020219275

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  • DOI: https://doi.org/10.1023/A:1007020219275

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