Abstract
The β-fructofuranosidase (Xd-INV) from the basidiomycota yeast Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma) is unique in its ability to synthesize neo- fructooligosaccharides (neo-FOS). In order to facilitate its industrial application, the recombinant enzyme expressed in Pichia pastoris (pXd-INV) was immobilized by entrapment in polyvinyl alcohol (PVA) hydrogels. The encapsulation efficiency exceeded 80%. The PVA lenticular particles of immobilized pXd-INV were stable up to approximately 40 °C. Using 600 g/L sucrose, the immobilized biocatalyst synthesized 18.9% (w/w) FOS (59.1 g/L of neokestose, 30.2 g/L of 1-kestose, 11.6 g/L of neonystose and 12.6 g/L of blastose). The operational stability of PVA-immobilized biocatalyst was assayed in a batch reactor at 30 °C. The enzyme preserved its initial activity during at least 7 cycles of 26 h.
Highlights
Fructooligosaccharides (FOS) are fructose oligomers linked to a sucrose skeleton by different β(2→1) or β(2→6) glycosidic bonds [1]
The β-fructofuranosidase Xd-INV from the yeast Xanthophyllomyces dendrorhous is a dimeric glycoprotein with a molecular mass of 320–380 kDa, which belongs to the glycoside hydrolase (GH) family 32 [9,10]
We have investigated the immobilization of recombinant pXd-INV to facilitate its industrial application in the production of neo- fructooligosaccharides (neo-FOS) and other fructosylated derivatives
Summary
Fructooligosaccharides (FOS) are fructose oligomers linked to a sucrose skeleton by different β(2→1) or β(2→6) glycosidic bonds [1]. In addition to their prebiotic properties—which promote the development of bifidobacteria and lactobacillus in the gastrointestinal tract [2]—and their low glycemic index, FOS may exert other benefits in human health, including a better gut absorption of. Commercial FOS possess an inulin-type structure containing β(2→1) linked fructose units [6]. It has been reported that neo-FOS, in which one fructosyl moiety is β(2→6) linked to the glucose unit of sucrose, could display improved prebiotic and physicochemical properties with regard to inulin-type FOS [7,8].
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