Production of Lacto-<i>N</i>-biose I Using Crude Extracts of Bifidobacterial Cells

DOI Web Site 参考文献22件 オープンアクセス
  • Machida Shuntaro
    Institute of Food Research, National Agriculture and Food Research Organization Present address: National Institute of Technology, Ube College
  • Saito Katsuichi
    Institute of Food Research, National Agriculture and Food Research Organization The late
  • Nishimoto Mamoru
    Institute of Food Research, National Agriculture and Food Research Organization
  • Kitaoka Motomitsu
    Institute of Food Research, National Agriculture and Food Research Organization Faculty of Agriculture, Niigata University

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<p>Lacto-N-biose I (LNB) is supposed to represent the bifidus factor in human milk oligosaccharides, and can be practically produced from sucrose and GlcNAc using four bifidobacterial enzymes, 1,3-β-galactosyl-N-acetylhexosamine phosphorylase, sucrose phosphorylase, UDP-glucose-hexose 1-phosphate uridylyltransferase, and UDP-glucose 4-epimerase, recombinantly produced by Escherichia coli. Here the production of LNB by the same enzymatic method without using genetically modified enzymes to consider the use of LNB for a food ingredient was reported. All four enzymes were produced as the intracellular enzymes of Bifidobacterium strains. The mixture of the crude extracts contained all four enzymes, with other enzymes interfering with the LNB production, namely, phosphoglucomutase, fructose 6-phosphate phosphoketolase, and glycogen phosphorylase. The first two interfering enzymes were selectively inactivated by heat treatment at 47 °C for 1 h in the presence of pancreatin, and glycogen phosphorylase was disabled by hydrolyzing its possible acceptor molecules using glucoamylase. Finally, 91 % of GlcNAc was converted into LNB in the 100-mL reaction mixture containing 300 mM GlcNAc.</p>

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