title: Synthesis and structural characterization of raffinosyl-oligofructosides upon transfructosylation by Lactobacillus gasseri DSM 20604 inulosucrase.
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.
authors.affiliation: Instituto de Investigación en Ciencias de la Alimentación, CIAL (CSIC-UAM), CEI (UAM+CSIC), C/ Nicolás Cabrera 9, 28049, Madrid, Spain.
authors.forename: Marina
authors.initials: M
authors.lastname: Díez-Municio
authors.affiliation: Instituto de Investigación en Ciencias de la Alimentación, CIAL (CSIC-UAM), CEI (UAM+CSIC), C/ Nicolás Cabrera 9, 28049, Madrid, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Herrero
authors.affiliation: Instituto de Ciencia y Tecnología de Alimentos y Nutrición, ICTAN (CSIC), C/ Juan de la Cierva 3, 28006, Madrid, Spain.
authors.forename: Blanca
authors.initials: B
authors.lastname: de Las Rivas
authors.affiliation: Instituto de Ciencia y Tecnología de Alimentos y Nutrición, ICTAN (CSIC), C/ Juan de la Cierva 3, 28006, Madrid, Spain.
authors.forename: Rosario
authors.initials: R
authors.lastname: Muñoz
authors.affiliation: Centro Química Orgánica "Lora-Tamayo" (CSIC), C/ Juan de la Cierva 3, 28006, Madrid, Spain.
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Jimeno
authors.affiliation: Instituto de Investigación en Ciencias de la Alimentación, CIAL (CSIC-UAM), CEI (UAM+CSIC), C/ Nicolás Cabrera 9, 28049, Madrid, Spain. javier.moreno@csic.es.
authors.forename: F Javier
authors.initials: FJ
authors.lastname: Moreno
mesh_terms: D003470:Culture Media / Q000737:chemistry; D005630:Fructans / Q000737:chemistry; D005632:Fructose / Q000737:chemistry; D006602:Hexosyltransferases / Q000378:metabolism*; D007218:Industrial Microbiology; D000069998:Lactobacillus gasseri / Q000201:enzymology*; D008279:Magnetic Resonance Imaging; D008553:Melibiose / Q000737:chemistry; D011887:Raffinose / Q000737:chemistry*; D011994:Recombinant Proteins / Q000378:metabolism; D013395:Sucrose / Q000737:chemistry; D001426:Bacterial Proteins
chemical_list: D003470:Culture Media; D005630:Fructans; D005632:Fructose; D006602:Hexosyltransferases; D008553:Melibiose; D011887:Raffinose; D011994:Recombinant Proteins; D013395:Sucrose; C021451:inulosucrase; D001426:Bacterial Proteins
keywords: Bioactive oligosaccharides; Inulosucrase; Raffinose; Transfructosylation reaction; α-Galactosides