Human desaturase gene and uses thereof

• US 6,432,684 B1
• Filed: 01/08/1999
• Issued: 08/13/2002
• Est. Priority Date: 04/11/1997
• Status: Expired due to Term

• Current Assignee: Abbott Laboratories Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Das, Tapas, Mukerji, Pradip, Huang, Yung-Sheng, Leonard, Amanda Eun-Yeong
• Primary Examiner(s): Saidha, Tekchand
• Application Number:
  US09/227,613
• Time in Patent Office: 1,313 Days
• Field of Search: 435/189, 435/136, 435/252.3, 435/320.1, 536/23.2, 530/350
• US Class Current: 435/136
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23L 33/10 using additives addition of substantially indigestible substances A23L33/21
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A23L 33/13 Nucleic acids or derivatives thereof A23L33/145 takes precedence
• CPC Class Codes: A23L 33/17 Amino acids, peptides or proteins
• CPC Class Codes: A23L 33/40 Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
• CPC Class Codes: A23L 7/126 Snacks or the like obtained by binding, shaping or compacting together cereal grains or cereal pieces, e.g. cereal bars
• CPC Class Codes: A23L 9/10 Puddings; Dry powder puddings
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A23V 2250/1862 Arachidonic acid
• CPC Class Codes: A23V 2250/1882 Polyunsaturated fatty acids
• CPC Class Codes: A23V 2250/19 Genetically modified oil
• CPC Class Codes: A61K 31/202 having three or more double bonds, e.g. linolenic eicosanoids, e.g. leukotrienes A61K31/557
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 8/361 Carboxylic acids having more than seven carbon atoms in an unbroken chain; Salts or anhydrides thereof
• CPC Class Codes: A61K 8/67 Vitamins
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: C12N 15/81 : for yeasts
• CPC Class Codes: C12N 15/8247 : involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0083 : Miscellaneous (1.14.99)
• 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 7/6472 : 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 a gene involved in the desaturation of polyunsaturated fatty acids at carbon 5 (i.e., “human Δ5-desaturase”) and to uses thereof. In particular, human Δ5-desaturase may be utilized, for example, in the conversion of dihomo-γ-linolenic acid (DGLA) to arachidonic acid (AA) and in the conversion of 20:4n-3 to eicosapentaenoic acid (EPA). AA or polyunsaturated fatty acids produced therefrom may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

Claims

1. An isolated, human nucleotide sequence corresponding to or complementary to at least about 70% of the nucleotide sequence comprising SEQ ID NO:

1, wherein said isolated nucleotide sequence encodes a polypeptide comprising SEQ ID NO;

12, which desaturates polyunsaturated fatty acids at carbon 5.

2. A method of producing a human Δ

5-desaturase comprising the steps of;

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

1 (FIG. 12);

b) constructing a vector comprising;

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

c) introducing said vector into a host cell under time and conditions sufficient for expression of said human Δ

5-desaturase.

3. The method of claim 2 wherein said host cell is a eukaryotic cell or a prokaryotic cell.

4. The method of claim 3 wherein said prokaryotic cell is selected from the group consisting of E. coli, cyanobacteria, and B. subtilis.

5. The method of claim 3 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

6. The method of claim 5 wherein said fungal cell is a yeast cell.

7. The method of claim 6 wherein said yeast cell is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Candida Spp., Lipomyces starkey, Yarrowia lipolytica, Kluyveromyces Spp., Hansenula spp., Trichoderma spp. and Pichia spp.

8. The method of claim 7 wherein said yeast cell is Saccharomyces cerevisiae.

9. A vector comprising:

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

1 (FIG.

     12) operably linked to

b) a promoter.

10. A host cell comprising said vector of claim 9.

11. The host cell of claim 10, wherein said host cell is a eukaryotic cell or a prokaryotic cell.

12. The host cell of claim 11 wherein said prokaryotic cell is selected from the group consisting of E. coli, Cyanobacteria, and B. subtilis.

13. The host cell of claim 11 wherein said eukaryotic cell is selected from the group consisting of a mammalian cell, an insect cell, a plant cell and a fungal cell.

14. The host cell of claim 13 wherein said fungal cell is a yeast cell.

15. The host cell of claim 14 wherein said yeast cell is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Candida spp., Lipomyces starkey, Yarrowia lipolytica, Kluyveromyces spp., Hansenula spp., Trichoderma spp. and Pichia spp.

16. The host cell of claim 15 wherein said host cell is Saccharomyces cerevisiae.

17. A mammalian cell comprising said vector of claim 9, wherein expression of said nucleotide sequence of said vector results in production of altered levels of arachidonic acid (AA) or eicosapentaenoic acid (EPA) when said cell is grown in a culture media comprising a fatty acid selected from the group consisting of an essential fatty acid, linoleic acid (LA) and α

-linolenic acid (ALA).