Fructose-4-epimerase and method of preparing tagatose using the same

Patent metadata

Field	Value
Publication number	US-11459594-B2
Application number	US-201917280854-A
Country	US
Kind code	B2
Filing date	Sep 27, 2019
Priority date	Sep 28, 2018
Publication date	Oct 4, 2022
Grant date	Oct 4, 2022

Abstract

Provided are a fructose-4-epimerase variant having tagatose conversion activity, and a method of producing tagatose using the same.

First claim

The invention claimed is:

1. A fructose-4-epimerase variant comprising substitution of another amino acid for a threonine (T) residue at position 124 from the N-terminus of fructose-4-epimerase including an amino acid sequence of SEQ ID NO: 1, wherein the variant includes an amino acid sequence having at least 60% sequence identity to the amino acid sequence of SEQ ID NO: 1.

2. The fructose-4-epimerase variant of claim 1 , wherein the another amino acid is a polar amino acid or a nonpolar amino acid, except threonine (T).

3. The fructose-4-epimerase variant of claim 1 , wherein the variant has further substitution of another amino acid for an amino acid at position 390.

4. The fructose-4-epimerase variant of claim 3 , wherein the substituted amino acid at position 390 is a nonpolar amino acid, an acidic amino acid, or a basic amino acid.

5. The fructose-4-epimerase variant of claim 1 , wherein the variant has further substitution of another amino acid for an amino acid at position 97.

6. The fructose-4-epimerase variant of claim 5 , wherein the another amino acid is tyrosine (Y).

7. The fructose-4-epimerase variant of claim 5 , wherein the variant has further substitution of another amino acid for any one of amino acids at positions 33, 80, 102, 137, 210, and 318.

8. The fructose-4-epimerase variant of claim 7 , wherein the another amino acid is selected from the group consisting of a polar amino acid, a nonpolar amino acid, an acidic amino acid, and a basic amino acid.

9. The fructose-4-epimerase variant of claim 1 , wherein the variant has further substitution of another amino acid for an amino acid at position 97 and substitution of another amino acid for an amino acid at position 367.

10. The fructose-4-epimerase variant of claim 9 , wherein the another amino acid at position 97 is tyrosine (Y) and another amino acid at position 367 is valine (V).

11. The fructose-4-epimerase variant of claim 9 , wherein the variant has further substitution of another amino acid for any one of amino acids at positions 102, 137, 210, 239, and 318.

12. The fructose-4-epimerase variant of claim 11 , wherein the another amino acid is selected from the group consisting of a nonpolar amino acid, a polar amino acid, and a basic amino acid.

13. The fructose-4-epimerase variant of claim 9 , wherein the variant has further substitution of another amino acid for an amino acid at position 33.

14. The fructose-4-epimerase variant of claim 13 , wherein the another amino acid is arginine (R).

15. The fructose-4-epimerase variant of claim 13 , wherein the variant has further substitution of another amino acid for any one of amino acids at positions 80, 102, 210, and 318.

16. The fructose-4-epimerase variant of claim 15 , wherein the another amino acid is selected from the group consisting of a basic amino acid, a nonpolar amino acid, and a polar amino acid.

17. The fructose-4-epimerase variant of claim 9 , wherein the variant has further substitution of another amino acid for an amino acid at position 80.

18. The fructose-4-epimerase variant of claim 17 , wherein the another amino acid is arginine (R).

19. The fructose-4-epimerase variant of claim 17 , wherein the variant has further substitution of another amino acid for any one of amino acids at positions 102, 137, and 210.

20. The fructose-4-epimerase variant of claim 19 , wherein the another amino acid is a nonpolar amino acid or a polar amino acid.

21. The fructose-4-epimerase variant of claim 9 , wherein the variant has further substitution of another amino acid for an amino acid at position 210 and substitution of another amino acid for an amino acid at position 318.

22. The fructose-4-epimerase variant of claim 21 , wherein the another amino acid at position 210 is serine (S) and another amino acid at position 318 is glycine (G).

23. A polynucleotide encoding the fructose-4-epimerase variant of claim 1 .

24. A vector comprising the polynucleotide of claim 23 .

25. A microorganism comprising the fructose-4-epimerase variant of claim 1 ; a polynucleotide encoding the fructose-4-epimerase variant; or a vector including the polynucleotide.

26. A composition for producing tagatose, the composition comprising the fructose-4-epimerase variant of claim 1 ; a microorganism expressing the fructose-4-epimerase variant; or a culture of the microorganism.

27. The composition for producing tagatose of claim 26 , the composition further comprising fructose.

28. A method of producing tagatose, the method comprising the step of converting fructose into tagatose by contacting fructose with the fructose-4-epimerase variant of claim 1 ; the microorganism including the fructose-4-epimerase variant; or the culture thereof.

Assignees

• Cj Cheiljedang Corp

Inventors

• Lee Young Mi
• Park Eul Soo
• Park Il Hyang
• Shin Sun Mi
• Yang Sung Jae
• Yoon Ran Young
• Choi Eun Jung
• Kim Seong Bo
• Park Seung Won

Classifications

• C12Y501/03

  acting on carbohydrates and derivatives (5.1.3) · CPC title

• C12P19/24 Primary

  produced by the action of an isomerase, e.g. fructose · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N9/88

  Lyases (4.) · CPC title