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Synthesis of α-galactooligosaccharides with α-galactosidase from Lactobacillus reuteri of canine origin

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Abstract

Crude cell-free extracts from Lactobacillus reuteri grown on cellobiose, maltose, lactose and raffinose were assayed for glycosidic activities. When raffinose was used as the carbon source, α-galactosidase was produced, showing the highest yield at the beginning of the stationary growth phase. A 64 kDa enzyme was purified by ultra- and gel filtration, and characterized for its hydrolytic and synthetic activity. Highest hydrolytic activity was found at pH 5.0 at 50 °C (K M 0.55 mM, V max 0.80 μmol min−1 mg−1 of protein). The crude cell-free extract was further used in glycosyl transfer reactions to synthesize oligosaccharides from melibiose and raffinose. At a substrate concentration of 23% (w/v) oligosaccharide mixtures were formed with main products being a trisaccharide at 26% (w/w) yield from melibiose after 8 h and a tetrasaccharide at 18% (w/w) yield from raffinose after 7 h. Methylation analysis revealed the trisaccharide to be 6′ α-galactosyl melibiose and the tetrasaccharide to be stachyose. In both cases synthesis ceased when hydrolysis of the substrate reached 50%.

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Authors and Affiliations

  1. School of Food Biosciences, The University of Reading, Whiteknights, PO Box 226, Reading, RG6 6AP, UK

    G. Tzortzis, G. R. Gibson & R. A. Rastall

  2. Institute of Food Research, Colney Lane, Norwich, NR4 7UA, UK

    A. J. Jay

  3. Waltham Centre for Pet Nutrition, Waltham-on-the-Wolds, Melton Mowbray, Leicestershire, LE14 4RT, UK

    M. L. A. Baillon

Authors
  1. G. Tzortzis
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  2. A. J. Jay
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  3. M. L. A. Baillon
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  4. G. R. Gibson
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  5. R. A. Rastall
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Correspondence to R. A. Rastall.

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Tzortzis, G., Jay, A.J., Baillon, M.L.A. et al. Synthesis of α-galactooligosaccharides with α-galactosidase from Lactobacillus reuteri of canine origin. Appl Microbiol Biotechnol 63, 286–292 (2003). https://doi.org/10.1007/s00253-003-1426-0

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  • DOI: https://doi.org/10.1007/s00253-003-1426-0

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