The present invention provides nucleic acids, vectors, host cells and methods for production of fructosyltransferase from Aspergillus japonicus. The invention represents an advancement in the field of genetic engineering and provides methods for obtaining high yield of a novel recombinant fructosyltransferase encoded by ft gene of Aspergillus japonicus as a secreted protein.

The disclosure relates to methods and compositions for the production of fructans using sucrose:sucrose 1-fructosyl-transferase (1-SST), fructan:fructan 1-fructosyltransferase (1-FFT), and/or sucrose fructan-6-fructosyltransferase (6-SFT) enzymes.

Genetic constructs capable of manipulating fructan biosynthesis in photosynthetic cells of a plant, said genetic constructs include a promoter operatively linked to a nucleic acid encoding a bacterial FT enzyme. These constructs can be used in modification of fructan biosynthesis in plants and, more particularly, for manipulating fructan biosynthesis in photosynthetic cells. The constructs can also be used for increasing plant biomass and, more particularly, for enhancing biomass yield and/or yield stability, including shoot and/or root growth in a plant, and for enhancing the productivity of biochemical pathways.

A stable fructooligosaccharide (FOS) composition is produced from a raw material containing sucrose by enzymatic activity in aqueous mixture, chromatographically separating non-FOS carbohydrates from the aqueous mixture and evaporating the aqueous mixture to yield a syrupy FOS composition of at least 65 °Bx, wherein at least part or essentially all of the organic acids and ions is formed in situ.

The present invention discloses an industrial scale method to obtain 1-kestose by the use of a recombinant fructosyltransferase (FTF), isolated from Festuca arundinacea, expressed constitutively in a non-saccharolytic yeast. In this invention, the recombinant FTF type sucrose:sucrose 1-fructosyltransferase (1-SSTrec) is produced constitutively, stable and at high yield, both in the culture supernatant and in intact cells of the host Pichia pastoris. Hence, the invention additionally provides a method for 1-SST production at industrial scale. The recombinant enzyme is then used for mass production of short-chain fructooligosaccharides (FOS), specifically 1-kestose, from sucrose. The method of the present invention establishes conditions that allow conversion rates where the synthesized FOS constitute above 55% (w/w) of the total sugars in the reaction mixture and the 1-kestose content reaches values higher than 90% (w/w) of the total FOS fraction.

The application discloses a method for producing an oligosaccharide of at least four monosaccharide units, advantageously an HMO, particularly an HMO of only four monosaccharide units, said method comprising a step of: culturing, in a culture medium containing a fucosylated, sialylated or N-acetyl-glucosaminylated lactose trisaccharide as acceptor, a genetically modified cell having a recombinant gene that encodes an enzyme capable of modifying said acceptor or one of the necessary intermediates in the biosynthetic pathway of the oligosaccharide of at least four monosaccharide units, advantageously an HMO, particularly an HMO of only four monosaccharide units, from said acceptor.

A process for the recovery of betaine from a molasses comprises a conversion step, in which the molasses is subjected to the action of an enzyme having endo-inulinase activity and/or fructosyltransferase activity, to form a fructan-containing molasses (fructan-molasses); a separation step, in which the fructan-molasses is subjected to a chromatographic separation, thereby obtaining a betaine-containing fraction.

The present invention discloses an industrial scale method to obtain 1-kestose by the use of a recombinant fructosyltransferase (FTF), isolated from Festuca arundinacea, expressed constitutively in a non-saccharolytic yeast. In this invention, the recombinant FTF type sucrose:sucrose 1-fructosyltransferase (1-SSTrec) is produced constitutively, stable and at high yield, both in the culture supernatant and in intact cells of the host Pichia pastoris. Hence, the invention additionally provides a method for 1-SST production at industrial scale. The recombinant enzyme is then used for mass production of short-chain fructooligosaccharides (FOS), specifically 1-kestose, from sucrose. The method of the present invention establishes conditions that allow conversion rates where the synthesized FOS constitute above 55% (w/w) of the total sugars in the reaction mixture and the 1-kestose content reaches values higher than 90% (w/w) of the total FOS fraction.

The present invention discloses an industrial scale method to obtain 1-kestose by the use of a recombinant fructosyltransferase (FTF), isolated from Festuca arundinacea, expressed constitutively in a non-saccharolytic yeast. In this invention, the recombinant FTF type sucrose:sucrose 1-fructosyltransferase (1-SSTrec) is produced constitutively, stable and at high yield, both in the culture supernatant and in intact cells of the host Pichia pastoris. Hence, the invention additionally provides a method for 1-SST production at industrial scale. The recombinant enzyme is then used for mass production of short-chain fructooligosaccharides (FOS), specifically 1-kestose, from sucrose. The method of the present invention establishes conditions that allow conversion rates where the synthesized FOS constitute above 55% (w/w) of the total sugars in the reaction mixture and the 1-kestose content reaches values higher than 90% (w/w) of the total FOS fraction.