Method of generating a saccharide containing a galactose and a fructose moiety employing enzyme with transgalactosylating activity
AssigneeDupont Nutrition Biosciences Aps
InventorsThomas EISELE, Jacob Flyvholm CRAMER, Morten Krog LARSEN
Life of this patent
Estimating the term…
Abstract
A method of generating a saccharide, especially lactulose or lactosucrose, in the presence of a transgalactosylase enzyme, is described herein.
Claims47
A method of generating a saccharide containing a galactose moiety and a fructose moiety, wherein:
- (a) the galactose moiety is linked to the fructose moiety;
- or
- (b) the galactose moiety and the fructose moiety are separated by at least one monosaccharide moiety other than galactose or fructose;
- the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- the method being carried out at a pH of 5.5 to 9.5;
- provided that
- (i) when the galactose moiety is linked to the fructose moiety, the concentration of the first saccharide is less than 0.43 mol/L and the concentration of the second saccharide is less than 0.5 mol/L;
- and
- (ii) when the galactose moiety and the fructose moiety are separated by at least one monosaccharide moiety other than galactose or fructose, the concentration of the first saccharide and/or the concentration of the second saccharide is less than 0.5 mol/L.
A method of generating a saccharide containing a galactose moiety and a fructose moiety, wherein:
- (a) the galactose moiety is linked to the fructose moiety;
- or
- (b) the galactose moiety and the fructose moiety are separated by at least one monosaccharide moiety other than galactose or fructose;
- the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- wherein the enzyme is selected from the group consisting of:
- a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO:
- 1, wherein said polypeptide consists of at most 980 amino acid residues;
- b) a polypeptide encoded by a polynucleotide that hybridizes under at least low stringency conditions with:
- i) the nucleic acid sequence comprised in SEQ ID NO:
- 9, encoding the polypeptide of SEQ ID NO:
- 1;
- or
- ii) the complementary strand of i).
A method of generating a saccharide in which a galactose moiety is linked to a fructose moiety, the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- the method being carried out at a pH of 5.5 to 9.5;
- wherein:
- the concentration of the first saccharide is less than 0.43 mol/L;
- and
- the concentration of the second saccharide is less than 0.8 mol/L.
The method of any preceding claim, wherein the enzyme is a β-galactosidase.
The method of any preceding claim, wherein the enzyme is classified in Enzyme Classification (E.C.) 3.2.1.23.
The method of any preceding claim, wherein the enzyme is of bacterial origin or fungal origin.
The method of any preceding claim, wherein the enzyme is selected from the group consisting of:
- a) a polypeptide having transgalactosylating activity and comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO:
- 1, wherein said polypeptide consists of at most 980 amino acid residues;
- b) a polypeptide encoded by a polynucleotide that hybridizes under at least low stringency conditions with:
- i) the nucleic acid sequence comprised in SEQ ID NO:
- 9, encoding the polypeptide of SEQ ID NO:
- 1;
- or
- ii) the complementary strand of i).
A method of generating a saccharide in which a galactose moiety is linked to a fructose moiety, the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- wherein the enzyme is selected from the group consisting of:
- a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO:
- 1, wherein said polypeptide consists of at most 980 amino acid residues;
- b) a polypeptide encoded by a polynucleotide that hybridizes under at least low stringency conditions with:
- i) the nucleic acid sequence comprised in SEQ ID NO:
- 9, encoding the polypeptide of SEQ ID NO:
- 1;
- or
- ii) the complementary strand of i).
The method of any preceding claim, carried out in situ in a food composition.
The method of any preceding claim, wherein the yield of lactulose is at least 10%, such as at least 12%, such as at least 15%, such as at least 18%, such as at least 20%, such as at least 22%, such as at least 25%, calculated by weight based on the total weight of lactose and fructose used as starting material.
A lactulose-containing composition obtainable by the method of any preceding claim.
A method of generating a saccharide containing a galactose moiety and a fructose moiety, the galactose moiety and the fructose moiety being separated by at least one monosaccharide moiety other than galactose or fructose, the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- the method being carried out at a pH of 5.5 to 9.5;
- wherein the concentration of the first saccharide and/or the concentration of the second saccharide is less than 0.5 mol/L.
A method of generating a saccharide containing a galactose moiety and a fructose moiety, the galactose moiety and the fructose moiety being separated by at least one monosaccharide moiety other than galactose or fructose, the method comprising:
- contacting a first saccharide, said first saccharide containing a galactose moiety, with a second saccharide, said second saccharide containing a fructose moiety, the first and the second saccharides being different,
- in the presence of an enzyme capable of catalysing the transfer of a galactose moiety to the second saccharide containing the fructose moiety,
- wherein the enzyme is selected from the group consisting of:
- a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO:
- 1, wherein said polypeptide consists of at most 980 amino acid residues;
- b) a polypeptide encoded by a polynucleotide that hybridizes under at least low stringency conditions with:
- i) the nucleic acid sequence comprised in SEQ ID NO:
- 9, encoding the polypeptide of SEQ ID NO:
- 1;
- or
- ii) the complementary strand of i).
A lactosucrose-containing composition obtainable by the method of any preceding claim.
Dependencies are inferred from the claim text.