Characterization of a beta-fructofuranosidase from Schwanniomyces occidentalis with transfructosylating activity yielding the prebiotic 6-kestose.
beta-Fructofuranosidases are powerful tools in industrial biotechnology. We have characterized an extracellular beta-fructofuranosidase from the yeast Schwanniomyces occidentalis. The enzyme shows broad substrate specificity, hydrolyzing sucrose, 1-kestose, nystose and raffinose, with different catalytic efficiencies (k(cat)/K(m)). Although the main reaction catalysed by this enzyme is sucrose hydrolysis, it also produces two fructooligosaccharides (FOS) by transfructosylation. A combination of (1)H, (13)C and 2D-NMR techniques shows that the major product is the prebiotic trisaccharide 6-kestose. The 6-kestose yield obtained with this beta-fructofuranosidase is, to our concern, higher than those reported with other 6-kestose-producing enzymes, both at the kinetic maximum (76gl(-1)) and at reaction equilibrium (44gl(-1)). The total FOS production in the kinetic maximum was 101gl(-1), which corresponded to 16.4% (w/w) referred to the total carbohydrates in the reaction mixture.
Alvaro-Benito
Miguel
M
Departamento de Biología Molecular (CSIC-UAM), Universidad Autónoma Madrid, Cantoblanco, 28049 Madrid, Spain.
de Abreu
Miguel
M
Fernández-Arrojo
Lucía
L
Plou
Francisco J
FJ
Jiménez-Barbero
Jesús
J
Ballesteros
Antonio
A
Polaina
Julio
J
Fernández-Lobato
María
M
D001709:Biotechnology; D007700:Kinetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry; D004718:Saccharomycetales / Q000201:enzymology*; D013379:Substrate Specificity; D014312:Trisaccharides / Q000096:biosynthesis* / Q000737:chemistry; D043324:beta-Fructofuranosidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D009844:Oligosaccharides; D014312:Trisaccharides; C116580:fructooligosaccharide; C073432:6-kestose; D043324:beta-Fructofuranosidase