title: An improved method for the production of fructooligosaccharides by immobilized β-fructofuranosidase from Sclerotinia sclerotiorum.
abstract: This work is focused on the prebiotic synthesis by a purified immobilized β-fructofuranosidase (FFase) using a by-product molasses as a substrate. When cultivated on wheat bran, the fungus Sclerotinia sclerotiorum produces FFase with interesting transfructosylating activity. The enzyme was purified by gel filtration and anion exchange chromatography to homogeneity. It showed a specific activity of 66.06 U/mg and a molecular mass of 50 kDa. The FFase was immobilized covalently on alginate and chitosan, and the immobilization yield was 90% and 81% respectively, yet the immobilization efficiency was 52% and 93% in that order. The fixed enzymes were stable at a pH varying from 4.0 to 7.0 and at a temperature ranging from 4 to 70 °C. Yet, kinetic parameters and catalytic efficiency were determined for both immobilized and free FFases. Interestingly, chitosan cross-linked enzyme activity was maintained at 89.24% level after 50 reuses during 1 week. Continuous production of fructooligosaccharides (FOS) from beet molasses in chitosan enzyme reactor was improved. The maximum production yield obtained in 12 H was 72.2% (g FOS/g Sucrose). Thin-layer chromatography analysis showed that the major products are kestose and nystose.
authors: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, Tunis Cedex, Tunisia.
authors: Refka
authors: R
authors: Mouelhi
authors: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, Tunis Cedex, Tunisia.
authors: Ferid
authors: F
authors: Abidi
authors: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, Tunis Cedex, Tunisia.
authors: Mohamed Nejib
authors: MN
authors: Marzouki
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D002855:Chromatography, Thin Layer; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry; D013696:Temperature; D043324:beta-Fructofuranosidase / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D009844:Oligosaccharides; C116580:fructooligosaccharide; D043324:beta-Fructofuranosidase
keywords: fructooligosaccharides; immobilization; molasses; purification; β-fructofuranosidase