title: Exploring the transferase activity of Ffase from Schwanniomyces occidentalis, a β-fructofuranosidase showing high fructosyl-acceptor promiscuity.
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.
authors.affiliation: Centro de Biología Molecular Severo Ochoa (CBMSO; CSIC-UAM), Departamento de Biología Molecular, Universidad Autónoma de Madrid, Nicolás Cabrera 1, 28049, Madrid, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Piedrabuena
authors.affiliation: Instituto de Catálisis y Petroleoquímica, CSIC, Marie Curie 2, 28049, Madrid, Spain.
authors.forename: Noa
authors.initials: N
authors.lastname: Míguez
authors.affiliation: 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.
authors.forename: Ana
authors.initials: A
authors.lastname: Poveda
authors.affiliation: Instituto de Catálisis y Petroleoquímica, CSIC, Marie Curie 2, 28049, Madrid, Spain.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Plou
authors.affiliation: Centro de Biología Molecular Severo Ochoa (CBMSO; CSIC-UAM), Departamento de Biología Molecular, Universidad Autónoma de Madrid, Nicolás Cabrera 1, 28049, Madrid, Spain. mfernandez@cbm.csic.es.
authors.forename: María
authors.initials: M
authors.lastname: Fernández-Lobato
mesh_terms: D007700:Kinetics; D004718:Saccharomycetales / Q000201:enzymology*; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism; D013402:Sugar Alcohols / Q000378:metabolism*; D043324:beta-Fructofuranosidase / Q000378:metabolism*
chemical_list: D013402:Sugar Alcohols; D013395:Sucrose; D043324:beta-Fructofuranosidase
keywords: Fructooligosaccharides; Fructosyl-alditols; Kinetic analysis; Schwanniomyces occidentalis; β-fructofuranosidase