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1. (WO2008064025) CLONAGE ET SÉQUENÇAGE DU GÈNE DE LA FÉRULATE ESTÉRASE À PARTIR DE <I>LACTOBACILLUS BUCHNERI</I> LN4017
Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

THAT WHICH IS CLAIMED:
1. A DNA construct comprising a promoter that drives expression in a plant or plant cell operably linked to a nucleotide sequence that encodes a polypeptide having ferulate esterase activity, wherein the nucleotide sequence is selected from the group consisting of:
a) a polynucleotide comprising SEQ ID NO: 1 , 2 or 4;
b) a polynucleotide having at least about 85% sequence identity to SEQ ID NO: 1 , 2 or 4;
c) a polynucleotide encoding SEQ ID NO: 3 or 5; and
d) a polynucleotide encoding a polypeptide having at least 85% sequence identity to SEQ ID NO: 3 or 5.

2. The DNA construct of claim 1 , wherein the nucleotide sequence is derived from a Lactobacillus species.

3. The DNA construct of claim 2, wherein the Lactobacillus species is Lactobacillus buchneri.

4. The DNA construct of claim 1 further comprising an operably linked polynucleotide encoding a signal peptide.

5. The DNA construct of claim 1 , wherein the DNA construct further comprises a polynucleotide sequence that targets expression of the
polynucleotide.

6. A transformed plant cell comprising at least one DNA construct according to claim 1.

7. The plant cell of claim 6, wherein said plant cell is from a monocot.

8. The plant cell of claim 7, wherein said monocot is a member of a genus selected from the group consisting of Festuca, Lolium, Sorghum, Zea, Thticum, Avena, and Poa.

9. A transgenic plant comprising at least one DNA construct according to claim 1.

10. The transgenic plant of claim 9, wherein said plant displays increased digestibility.

11. The transgenic plant of claim 9, wherein the nucleotide sequence is derived from a Lactobacillus species.

12. The transgenic plant of claim 11 , wherein the Lactobacillus species is Lactobacillus buchneri.

13. The transgenic plant of claim 9 wherein the promoter is selected from the group consisting of:
a) an inducible promoter;
b) a senescence promoter;
c) an embryo specific promoter;
d) a pericarp specific promoter;
e) a heat shock promoter; and
f) a constitutive promoter.

14. The transgenic plant of claim 9, wherein said plant is a monocot.

15. The transgenic plant of claim 14, wherein said monocot is a member of a genus selected from the group consisting of Festuca, Lolium, Sorghum, Zea,

Thticum, Avena, and Poa.

16. A transformed seed of the transgenic plant of claim 9, wherein the seed comprises the construct.

17. The transgenic plant of claim 9, further comprising introduction into the plant a second DNA construct comprising a promoter operably linked to a xylanase encoding polynucleotide.

18. The transgenic plant of claim 17, wherein the first and second DNA constructs are present on separate plasmids.

19. A method of affecting the cross-linking of phenolic acids in plant cell walls of a transgenic plant, the method comprising introducing into the plant a

DNA construct comprising a promoter operably linked to an isolated
polynucleotide encoding a ferulate esterase, wherein said polynucleotide has a sequence selected from the group consisting of:
a) a polynucleotide comprising SEQ ID NO: 1 , 2 or 4;
b) a polynucleotide having at least about 85% sequence identity to SEQ ID NO: 1 , 2 or 4;
c) a polynucleotide encoding SEQ ID NO: 3 or 5; and
d) a polynucleotide encoding a polypeptide having at least 85% sequence identity to SEQ ID NO: 3 or 5.

20. The method of claim 19, wherein the polynucleotide is derived from a Lactobacillus species.

21. The method of claim 20, wherein the Lactobacillus species is
Lactobacillus buchneri.

22. The method of claim 19, wherein said plant is a monocot.

23. The method of claim 22, wherein said monocot is a member of a genus selected from the group consisting of Festuca, Lolium, Sorghum, Zea,

Thticum, Avena, and Poa.

24. The method of claim 19, further comprising introduction into the plant a second DNA construct comprising a promoter operably linked to a xylanase encoding polynucleotide.

25. The method of claim 24, wherein the first and second DNA
constructs are present on separate plasmids.

26. A transgenic plant produced by the method of claim 19.

27. A method for increasing digestibility of a plant or plant part fed to an animal, the method comprising introducing into a plant a DNA construct
comprising a promoter operably linked to a polynucleotide encoding ferulate esterase, wherein said polynucleotide has a sequence selected from the group consisting of:
i) a polynucleotide comprising SEQ ID NO: 1 or 2;
ii) a polynucleotide having at least about 85% sequence identity to SEQ ID NO: 1 , 2 or 4;
iii) a polynucleotide encoding SEQ ID NO: 3 or 5; and
iv) a polynucleotide encoding a polypeptide having at least 85% sequence identity to SEQ ID NO: 3 or 5.

28. The method of claim 27, wherein the polynucleotide is derived from

Lactobacillus buchneri.

29. The method of claim 27, wherein said plant is a monocot.

30. The method of claim 29, wherein said monocot is a member of a genus selected from the group consisting of Festuca, Lolium, Sorghum, Zea, Thticum, Avena, and Poa.

31. The method of claim 27, further comprising introduction into the plant a second DNA construct comprising a promoter operably linked to a xylanase encoding polynucleotide.

32. The method of claim 31 , wherein the first and second DNA
constructs are present on separate plasmids.

33. A transgenic plant produced by the method of claim 27.

34. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NOs: 3 or 5; and
b) a polypeptide having at least 85% sequence identity to SEQ ID NOs: 3 or 5, wherein said polypeptide has ferulate esterase activity.

35. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:
a) a polynucleotide comprising the sequence set forth in SEQ ID NOs: 1 , 2 or 4;
b) a polynucleotide having at least about 85% sequence identity to SEQ ID NOs: 1 , 2 or 4;
c) a polynucleotide encoding the amino acid sequence of SEQ ID NOs: 3 or 5; and
d) a polynucleotide encoding the amino acid sequence of a polypeptide having at least 85% sequence identity to SEQ ID NOs: 3 or 5, wherein said polypeptide has ferulate esterase activity.

36. The nucleic acid molecule of claim 35, wherein said polypeptide has ferulate esterase activity.

37. The nucleic acid molecule of claim 35, wherein said nucleotide
sequence is optimized for expression in a plant.

38. A DNA construct comprising a promoter that drives expression in a plant or plant cell operably linked to a nucleotide sequence that encodes a polypeptide having acetyl esterase activity, wherein the nucleotide sequence is selected from the group consisting of:
a) a polynucleotide comprising SEQ ID NO: 4;
b) a polynucleotide having at least about 85% sequence identity to SEQ ID NO: 4;
c) a polynucleotide encoding SEQ ID NO: 5; and
d) a polynucleotide encoding a polypeptide having at least 85% sequence identity to SEQ ID NO: 5.

39. The DNA construct of claim 38, wherein the nucleotide sequence is derived from a Lactobacillus species.

40. The DNA construct of claim 39, wherein the Lactobacillus species is Lactobacillus buchneri.

41. A transformed plant cell comprising at least one DNA construct according to claim 38.

42. A transgenic plant comprising at least one DNA construct according to claim 38.

43. The transgenic plant of claim 42, wherein said plant displays increased digestibility.

44. The transgenic plant of claim 42, wherein said plant is a monocot.

45. The transgenic plant of claim 44, wherein said monocot is a member of a genus selected from the group consisting of Festuca, Lolium, Sorghum, Zea, Thticum, Avena, and Poa.