The ?-Fructofuranosidase from Rhodotorula dairenensis: Molecular Cloning, Heterologous Expression, and Evaluation of Its Transferase Activity

The ?-fructofuranosidase from the yeast Rhodotorula dairenensis (RdINV) produces a mixture of potential prebiotic fructooligosaccharides (FOS) levan-, inulin- and neo-FOS series by transfructosylation sucrose. In this work, gene responsible for activity was characterized its functionality proved in Pichia pastoris. amino acid sequence new protein contained most characteristic elements ?-fructofuranosidases included family 32 glycosyl hydrolases (GH32). heterologous produced about 170 kDa, where N-linked O-linked carbohydrates constituted 15% 38% total mass, respectively. Biochemical kinetic properties were similar to native enzyme, including ability produce sugars. maximum concentration FOS obtained 82.2 g/L, which 6-kestose represented 59% (w/w) products synthesized. RdINV fructosylate 19 hydroxylated compounds also explored, eight sugars four alditols modified. flexibility recognize diverse fructosyl acceptors makes valuable novel glycosyl-compounds with applications food pharmaceutical industries.