Glycosyltransferase glycosylating flavokermesic acid and/or kermesic acid
An isolated glycosyltransferase (GT) polypeptide capable of: (I): conjugating glucose to flavokermesic acid (FK); and/or (II): conjugating glucose to kermesic acid (KA) and use of this GT to e.g. make Carminic acid.
1. A method for producing flavokermesic acid glycoside or kermesic acid glycoside comprising the acts of:
providing an aglycon wherein said aglycon comprises flavokermesic acid or kermesic acid;
providing a sugar, wherein said sugar comprises a nucleotide activated glucose;
providing a recombinant cell wherein said recombinant cell comprises an expression cassette comprising an isolated polynucleotide comprising a nucleotide sequence which encodes an isolated glycosyltransferase polypeptide capable of:
(I): conjugating nucleotide activated glucose to flavokermesic acid (FK); and/or
(II): conjugating nucleotide activated glucose to kermesic acid (KA);
and wherein the glycosyltransferase polypeptide is at least one polypeptide selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence which has at least 95% identity with amino acids 1 to 515 of SEQ ID NO:2; and
(b) a polypeptide comprising an amino acid sequence which has at least 95% identity with amino acids 20 to 468 of SEQ ID NO:2; and
wherein said polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in an expression host; and
contacting said glycosyltransferase polypeptide with said aglycon and said nucleotide activated glucose to produce flavorkermesic acid glycoside or kermesic acid glycoside.
2. The method of claim 1 , wherein said cell is Saccharomyces spp. or Pichia spp.
3. The method of claim 2 , wherein said cell is Saccharomyces cerevisiae.
4. The method of claim 1 , wherein said nucleotide activated glucose comprises UDP-glucose.
5. The method of claim 1 , wherein said expression cassette comprises a plasmid.
6. The method of claim 1 , further comprising the act of purifying said produced flavokermesic acid glycoside or kermesic acid glycoside.
7. A method for producing flavokermesic acid glycoside or kermesic acid glycoside in vitro comprising the acts of:
providing an aglycon wherein said aglycon comprises flavokermesic acid or kermesic acid;
providing a sugar, wherein said sugar comprises a nucleotide activated glucose;
providing an isolated recombinant glycosyltransferase produced by recombinant cell wherein said recombinant cell comprises an expression cassette comprising an isolated polynucleotide comprising a nucleotide sequence which encodes an isolated glycosyltransferase polypeptide capable of:
(I): conjugating nucleotide activated glucose to flavokermesic acid (FK); and/or
(II): conjugating nucleotide activated glucose to kermesic acid (KA);
and wherein the glycosyltransferase polypeptide is at least one polypeptide selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence which has at least 95% identity with amino acids 1 to 515 of SEQ ID NO:2; and
(b) a polypeptide comprising an amino acid sequence which has at least 95% identity with amino acids 20 to 468 of SEQ ID NO:2; and
wherein said polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in an expression host; and
contacting said glycosyltransferase polypeptide with said aglycon and said nucleotide activated glucose to produce flavokermesic acid glycoside or kermesic acid glycoside.
8. The method of claim 7 , wherein said cell is Saccharomyces spp. or Pichia spp.
9. The method of claim 8 , wherein said cell is Saccharomyces cerevisiae.
10. The method of claim 7 , wherein said nucleotide activated glucose comprises UDP-glucose.
11. The method of claim 7 , wherein said isolated recombinant glycosyltransferase is isolated by the act of extraction from said recombinant cell comprising a expression cassette which encodes for a glycosyltransferase.