Elongase genes and uses thereof

• US 6,913,916 B1
• Filed: 07/24/2000
• Issued: 07/05/2005
• Est. Priority Date: 09/02/1998
• Status: Expired due to Fees

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Parker-Barnes, Jennifer M., Das, Tapas, Mukerji, Pradip, Huang, Yung-Sheng, Thurmond, Jennifer, Leonard, Amanda Eun-Yeong
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): Ramirez, Delia M.
• Application Number:
  US09/624,670
• Time in Patent Office: 1,807 Days
• Field of Search: 435/183, 435/4, 435/69.1, 435/325, 435/410, 435/252.3, 435/252.31, 435/252.33, 435/254.2, 435/254.21, 435/320.1, 435/419, 435/254.11, 435/254.22, 435/254.3, 435/254.5, 435/254.4, 435/254.7, 435/254.23, 435/348, 800/295, 536/23.2
• US Class Current: 435/183
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6431 Linoleic acids [18:2[n-6]]
• CPC Class Codes: C12P 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

The subject invention relates to the identification of several genes involved in the elongation of polyunsaturated acids (i.e., “elongases”) and to uses thereof. At least two of these genes are also involved in the elongation of monounsaturated fatty acids. In particular, elongase is utilized in the conversion of gamma linolenic acid (GLA) to dihomogama linolenic acid (DGLA) and in the conversion of DGLA or 20:4n-3 to eicosapentaenoic acid (EPA). DGLA may be utilized in the production of polyunsaturated fatty acids, such as arachiodonic acid (AA), docosahexaenoic acid (DHA), EPA, adrenic acid, ω6-docosapentaenoic acid or ω3-docosapentaenoic acid which may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

Claims

1. An isolated nucleic acid selected from the group consisting of:

(a) a nucleic acid comprising the polynucleotide of SEQ ID NO;

5 and (5) a nucleic acid completely complementary to the polynucleotide of (a).

2. The isolated nucleic acid of claim 1, wherein said isolated nucleic acid encodes a functionally active elongase which utilizes a polyunsaturated fatty acid as a substrate.

3. The isolated nucleic acid of claim 1, wherein said isolated nucleic acid is derived from a mammal.

4. The isolated nucleic acid of claim 3, wherein said isolated nucleic acid is derived from a mouse.

5. A method of producing an elongase enzyme comprising the steps of:

a) isolating a polynucleotide comprising the nucleotide sequence of SEQ ID NO;

5 or SEQ ID NO;

6;

b) constructing a vector comprising said polynucleotide of step (a) operably linked to a promoter; and

c) introducing said vector into a host cell for a time and under conditions sufficient for expression of said elongase enzyme.

6. The method of claim 5 wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

7. The method of claim 6 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

8. The method of claim 6 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

9. The method of claim 8 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

10. The method of claim 9 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

11. The method of claim 10 wherein said yeast cell is Saccharomyces cerevisiae.

12. A vector comprising a polynucleotide operably linked to a promoter, wherein said polynucleotide comprises the nucleotide sequence of SEQ ID NO:

5.

13. A host cell comprising the vector of claim 12.

14. The host cell of claim 13, wherein said host cell is selected from the group consisting of a eukaryotic cell or a prokaryotic cell.

15. The host cell of claim 14 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

16. The host cell of claim 14 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

17. The host cell of claim 16 wherein said fungal cell is selected from the group consisting of Saccharomyces spp., Candida spp., Lipomyces starkey, Yarrowia spp., Kluyveromyces spp., Hansenula spp., Aspergillus spp., Penicillium spp., Neurospora spp., Trichoderma spp. and Pichia spp.

18. The host cell of claim 17 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

19. The host cell of claim 18 wherein said host cell is Saccharomyces cerevisiae.

20. A plant cell, plant or plant tissue comprising the vector of claim 12, wherein expression of the polynucleotide of said vector results in production of a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

21. The plant cell, plant or plant tissue of claim 20, wherein said polyunsaturated fatty acid is selected from the group consisting of arachidonic acid (AA), adrenic acid (ADA), γ

-linoleic acid (GLA) and stearidonic acid (STA).

22. An isolated nucleic acid selected from the group consisting of:

(a) a nucleic acid comprising the polynucleotide of SEQ ID NO;

6 and (b) a nucleic acid completely complementary to the polynucleotide of (a).

23. An isolated polynucleotide encoding a polypeptide, wherein said polypeptide has elongase activity and comprises an amino acid sequence which is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:

63.

24. An isolated polynucleotide encoding a polypeptide, wherein said polypeptide has elongase activity and comprises an amino acid sequence which is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:

64.