Elongase genes and uses thereof

• US 7,070,970 B2
• Filed: 04/04/2003
• Issued: 07/04/2006
• Est. Priority Date: 08/23/1999
• Status: Expired due to Fees

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Parker-Barnes, Jennifer, Das, Tapas, Mukerji, Pradip, Huang, Yung-Sheng, Thurmond, Jennifer M., Leonard, Amanda E.
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): SWOPE, SHERIDAN
• Application Number:
  US10/408,736
• Publication Number:
  US 20030177508A1
• Time in Patent Office: 1,187 Days
• Field of Search: 435/193, 435/252.3, 435/252.33, 435/325, 435/320.1, 435/254.1, 435/254.2, 435/252.31, 435/348, 435/254.11, 435/419, 435/254.21, 435/254.22, 435/254.23, 435/254.3, 435/254.4, 435/254.5, 435/254.6, 536/23.2
• US Class Current: 435/193
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• 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 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]

Abstract

The subject invention relates to the identification of four genes involved in the elongation of polyunsaturated acids (i.e., “elongases”) and to uses thereof. 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 dihomogamma 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 arachidonic 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 molecule having a nucleotide sequence with at least 95% homology to SEQ ID NO:

2, or the complement thereof, wherein the nucleic acid sequence encodes a functionally active elongase that utilizes a polyunsaturated fatty acid as a substrate.

2. The isolated nucleic acid molecule of claim 1 wherein said sequence is represented by SEQ ID NO:

2.

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

a) isolating the nucleotide sequence represented by SEQ ID NO;

2;

b) constructing a vector comprising;

i) said isolated nucleotide sequence operably linked to ii) a promoter; and

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

4. The method of claim 3 wherein said host cell is selected from the group consisting of a eukaryotic cell and a prokaryotic cell.

5. The method of claim 4 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

6. The method of claim 4 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

7. The method of claim 6 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.

8. The method of claim 7 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

9. The method of claim 8 wherein said yeast cell is Saccharomyces cerevisiae.

10. A vector comprising:

a) a nucleotide sequence as represented by SEQ ID NO;

2 operably linked to b) a promoter.

11. A host cell comprising said vector of claim 10.

12. The host cell of claim 11, wherein said host cell is selected from the group consisting of a eukaryotic cell and a prokaryotic cell.

13. The host cell of claim 12 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

14. The host cell of claim 12 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

15. The host cell of claim 14 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.

16. The host cell of claim 15 wherein said fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Candida spp., Hansenula spp. and Pichia spp.

17. The host cell of claim 16 wherein said host cell is Saccharomyces cerevisiae.

18. A plant cell, plant or plant tissue comprising said vector of claim 10, wherein expression of said nucleotide sequence of said vector results in production of a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

19. The plant cell, plant or plant tissue of claim 18 wherein said polyunsaturated fatty acid is selected from the group consisting of DGLA, 20:

4n-3, and ADA.