title: Enzymatic synthesis and characterization of fructooligosaccharides and novel maltosylfructosides by inulosucrase from Lactobacillus gasseri DSM 20604.
abstract: The ability of an inulosucrase (IS) from Lactobacillus gasseri DSM 20604 to synthesize fructooligosaccharides (FOS) and maltosylfructosides (MFOS) in the presence of sucrose and sucrose-maltose mixtures was investigated after optimization of synthesis conditions, including enzyme concentration, temperature, pH, and reaction time. The maximum formation of FOS, which consist of β-2,1-linked fructose to sucrose, was 45% (in weight with respect to the initial amount of sucrose) and was obtained after 24 h of reaction at 55°C in the presence of sucrose (300 g liter(-1)) and 1.6 U ml(-1) of IS-25 mM sodium acetate buffer-1 mM CaCl2 (pH 5.2). The production of MFOS was also studied as a function of the initial ratios of sucrose to maltose (10:50, 20:40, 30:30, and 40:20, expressed in g 100 ml(-1)). The highest yield in total MFOS was attained after 24 to 32 h of reaction time and ranged from 13% (10:50 sucrose/maltose) to 52% (30:30 sucrose/maltose) in weight with respect to the initial amount of maltose. Nuclear magnetic resonance (NMR) structural characterization indicated that IS from L. gasseri specifically transferred fructose moieties of sucrose to either C-1 of the reducing end or C-6 of the nonreducing end of maltose. Thus, the trisaccharide erlose [α-d-glucopyranosyl-(1→4)-α-d-glucopyranosyl-(1→2)-β-d-fructofuranoside] was the main synthesized MFOS followed by neo-erlose [β-d-fructofuranosyl-(2→6)-α-d-glucopyranosyl-(1→4)-α-d-glucopyranose]. The formation of MFOS with a higher degree of polymerization was also demonstrated by the transfer of additional fructose residues to C-1 of either the β-2,1-linked fructose or the β-2,6-linked fructose to maltose, revealing the capacity of MFOS to serve as acceptors.
authors: Instituto de Investigación en Ciencias de la Alimentación, CIAL (CSIC-UAM), CEI (UAM+CSIC), Madrid, Spain.
authors: Marina
authors: M
authors: Díez-Municio
authors: Blanca
authors: B
authors: de las Rivas
authors: Maria Luisa
authors: ML
authors: Jimeno
authors: Rosario
authors: R
authors: Muñoz
authors: F Javier
authors: FJ
authors: Moreno
authors: Miguel
authors: M
authors: Herrero
mesh_terms: D019149:Bioreactors; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008401:Gas Chromatography-Mass Spectrometry; D006602:Hexosyltransferases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007778:Lactobacillus / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D008320:Maltose; D009844:Oligosaccharides / Q000096:biosynthesis*; D012394:Rosaniline Dyes; D013395:Sucrose; D013696:Temperature; D013997:Time Factors; D014312:Trisaccharides / Q000096:biosynthesis*; D001426:Bacterial Proteins
chemical_list: D017931:DNA Primers; D006602:Hexosyltransferases; D008320:Maltose; D009844:Oligosaccharides; D012394:Rosaniline Dyes; D013395:Sucrose; D014312:Trisaccharides; C116580:fructooligosaccharide; C043499:maltosylfructoside; C026440:glucosylsucrose; C048139:Coomassie blue; C021451:inulosucrase; D001426:Bacterial Proteins
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