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Exploring the transferase activity of Ffase from Schwanniomyces occidentalis, a β-fructofuranosidase showing high fructosyl-acceptor promiscuity

Applied Microbiology and Biotechnology volume 100, pages 8769–8778 (2016)Cite this article

Abstract

The β-fructofuranosidase from the yeast Schwanniomyces occidentalis (Ffase) produces the prebiotic sugars 6-kestose and 1-kestose by transfructosylation of sucrose, which makes it of biotechnological interest. In this study, the hydrolase and transferase activity of this enzyme was kinetically characterized and its potential to synthesize new fructosylated products explored. A total of 40 hydroxylated compounds were used as potential fructosyl-acceptor alternatives to sucrose. Only 17 of them, including some monosaccharides, disaccharides, and oligosaccharides as well as alditols and glycosides were fructosylated. The best alternative acceptors were the alditols. The major transfer product of the reaction including mannitol was purified and characterized as 1-O-β-d-fructofuranosyl-d-mannitol, whose maximum concentration reached 44 g/L, representing about 7.3 % of total compounds in the mixture and 89 % of all products generated by transfructosylation. The reactions including erythritol produced 35 g/L of an isomer mixture comprising 1- and 4-O-β-d-fructofuranosyl-d-erythritol. In addition, Ffase produced 24 g/L of the disaccharide blastose by direct fructosylation of glucose, which makes it the first enzyme characterized from yeast showing this ability. Thus, novel fructosylated compounds with potential applications in food and pharmaceutical industries can be obtained due to the Ffase fructosyl-acceptor promiscuity.

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Acknowledgments

This work was supported by the grant from the Spanish Ministry of Economy and Competitiveness: BIO2013-48779-C4-1/-4 and by an institutional grant from the Fundación Ramón Areces to the Centro de Biología Molecular Severo Ochoa. D.P. was supported by the Spanish Ministry of Education, Culture, and Sport fellowship ref. FPU014/01004. We thank Mrs. Asunción Martín-Redondo for the technical support.

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Affiliations

  1. Centro de Biología Molecular Severo Ochoa (CBMSO; CSIC-UAM), Departamento de Biología Molecular, Universidad Autónoma de Madrid, Nicolás Cabrera 1, 28049, Madrid, Spain

    David Piedrabuena & María Fernández-Lobato

  2. Instituto de Catálisis y Petroleoquímica, CSIC, Marie Curie 2, 28049, Madrid, Spain

    Noa Míguez & Francisco J. Plou

  3. CIC bioGUNE: Centre for Cooperative Research in Biosciences, Basque Network of Science, Technology and Innovation, Biscay Science and Technology Park, 801-A Ibaizabal Bidea St., Derio Town, 48160, Spain

    Ana Poveda

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  1. David Piedrabuena
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  2. Noa Míguez
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  3. Ana Poveda
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  4. Francisco J. Plou
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  5. María Fernández-Lobato
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Correspondence to María Fernández-Lobato.

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This study was funded by the Spanish Ministry of Economy and Competitiveness (BIO2013-48779-C4-1/-4).

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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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Piedrabuena, D., Míguez, N., Poveda, A. et al. Exploring the transferase activity of Ffase from Schwanniomyces occidentalis, a β-fructofuranosidase showing high fructosyl-acceptor promiscuity. Appl Microbiol Biotechnol 100, 8769–8778 (2016). https://doi.org/10.1007/s00253-016-7628-z

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  • DOI: https://doi.org/10.1007/s00253-016-7628-z

Keywords

  • β-fructofuranosidase
  • Schwanniomyces occidentalis
  • Fructooligosaccharides
  • Fructosyl-alditols
  • Kinetic analysis