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Synthesis and structural characterization of raffinosyl-oligofructosides upon transfructosylation by Lactobacillus gasseri DSM 20604 inulosucrase

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Abstract

A new process based on enzymatic synthesis of a series of raffinose-derived oligosaccharides or raffinosyl-oligofructosides (RFOS) with degree of polymerization (DP) from 4 to 8 was developed in the presence of raffinose. This process involves a transfructosylation reaction catalyzed by an inulosucrase from Lactobacillus gasseri DSM 20604 (IS). The main synthesized RFOS were structurally characterized by nuclear magnetic resonance (NMR). According to the elucidated structures, RFOS consist of β-2,1-linked fructose unit(s) to raffinose: α-d-galactopyranosyl-(1 → 6)-α-d-glucopyranosyl-(1↔2)-β-d-fructofuranosyl-((1 ← 2)-β-d-fructofuranoside)n (where n refers to the number of transferred fructose moieties). The maximum yield of RFOS was 33.4 % (in weight respect to the initial amount of raffinose) and was obtained at the time interval of 8–24 h of transfructosylation reaction initiated with 50 % (w/v) of raffinose. Results revealed the high acceptor and donor affinity of IS towards raffinose, being fairly comparable with that of sucrose for the production of fructooligosaccharides (FOS), including when both carbohydrates coexisted (sucrose/raffinose mixture, 250 g L−1 each). The production of RFOS was also attempted in the presence of sucrose/melibiose mixtures; in this case, the predominant acceptor-product formed was raffinose followed by a minor production of a series of oligosaccharides with varying DP. The easiness of RFOS synthesis and the structural similarities with both raffinose and fructan series of oligosaccharides warrant the further study of the potential bioactive properties of these unexplored oligosaccharides.

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Acknowledgments

M. Díez-Municio is supported by CSIC through JAE-Pre Programme co-funded by European Social Fund (ESF). M. Herrero thanks MICINN for his “Ramón y Cajal” contract. The authors gratefully acknowledge Dr. A. Gonçalves for the cloning of IS.

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Authors and Affiliations

  1. Instituto de Investigación en Ciencias de la Alimentación, CIAL (CSIC-UAM), CEI (UAM+CSIC), C/ Nicolás Cabrera 9, 28049, Madrid, Spain

    Marina Díez-Municio, Miguel Herrero & F. Javier Moreno

  2. Instituto de Ciencia y Tecnología de Alimentos y Nutrición, ICTAN (CSIC), C/ Juan de la Cierva 3, 28006, Madrid, Spain

    Blanca de las Rivas & Rosario Muñoz

  3. Centro Química Orgánica “Lora-Tamayo” (CSIC), C/ Juan de la Cierva 3, 28006, Madrid, Spain

    M. Luisa Jimeno

Authors
  1. Marina Díez-Municio
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  2. Miguel Herrero
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  3. Blanca de las Rivas
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  4. Rosario Muñoz
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  5. M. Luisa Jimeno
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  6. F. Javier Moreno
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Correspondence to F. Javier Moreno.

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Funding

This study was funded by the Ministerio de Economía y Competitividad (grant number AGL2011-27884) and by the Spanish Danone Institute.

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All authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Díez-Municio, M., Herrero, M., de las Rivas, B. et al. Synthesis and structural characterization of raffinosyl-oligofructosides upon transfructosylation by Lactobacillus gasseri DSM 20604 inulosucrase. Appl Microbiol Biotechnol 100, 6251–6263 (2016). https://doi.org/10.1007/s00253-016-7405-z

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