Δ4-desaturase genes and uses thereof

• US 7,045,683 B2
• Filed: 04/11/2002
• Issued: 05/16/2006
• Est. Priority Date: 05/04/2001
• Status: Expired due to Fees

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Das, Tapas, Mukerji, Pradip, Huang, Yung-Sheng, Thurmond, Jennifer, Pereira, Suzette L., Leonard, Amanda Eun-Yeong
• Primary Examiner(s): Nashed, Nashaat T.
• Application Number:
  US10/120,637
• Publication Number:
  US 20030134400A1
• Time in Patent Office: 1,496 Days
• Field of Search: 435/189, 435/252.33, 435/410, 435/320.1, 435/254.1, 435/253.33, 536/23.2, 536/23.74, 536/23.1, 800/298
• US Class Current: 800/298
• 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 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 9/12 Ready-to-eat liquid or semi-liquid desserts, e.g. puddings, not to be mixed with liquids, e.g. water, milk
• 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: A61P 3/02 Nutrients, e.g. vitamins, minerals
• 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 7/6434 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 genes involved in the desaturation of polyunsaturated fatty acids at carbon 4 (i.e., “Δ4-desaturase”). In particular, Δ4-desaturase may be utilized, for example, in the conversion of adrenic acid to ω6-docosapentaenoic acid and in the conversion of ω3-docosapentaenoic acid to docosahexaenoic acid. The polyunsaturated fatty acids produced by use of the enzyme may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

Claims

1. An isolated nucleic acid sequence or fragment thereof comprising or complementary to a nucleotide sequence encoding a polypeptide having Δ

4 fatty acid desaturase activity, wherein the amino acid sequence of said polypeptide has at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO;

37.

3. The isolated nucleic acid sequence of claim 1 or claim 2 wherein said sequence encodes a functionally active desaturase which utilizes a monounsaturated or polyunsaturated fatty acid as a substrate.

4. The isolated nucleic acid sequence of claim 1 or claim 2 wherein said sequence is derived from an organism selected from the group consisting of a fungus and an algae.

5. The isolated nucleic acid sequence of claim 4 wherein said sequence comprising SEQ ID NO:

36 is derived from the fungus Schizochytrium aggregatum.

2. An isolated nucleic acid sequence or fragment thereof comprising or complementary to a nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO:

36, and encodes Δ

4-fatty acid desaturase activity.

6. A method of producing a desaturase comprising the steps of:

a) isolating a nucleic acid sequence comprising or complementary to a nucleotide sequence;

i) encoding a polypeptide having an amino acid sequence having at least 95% amino acid identity to the amino acid sequence of SEQ ID NO;

37 and having Δ

4-fatty acid desaturase activity, or

ii) having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO;

36 and encoding Δ

4-fatty acid desaturase activity;

b) constructing a vector comprising;

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

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

7. A vector comprising:

a) an isolated nucleic acid sequence comprising or complementary to a nucleotide sequence;

i) encoding a polypeptide having an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence SEQ ID NO;

37 and Δ

4-fatty acid desaturase activity, or

ii) having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO ;

36 and encoding Δ

4-fatty acid desaturase activity, operably linked to

b) a promoter.

8. A host cell comprising said vector of claim 7.

9. A plant cell, plant or plant tissue comprising said vector of claim 7 wherein expression of said nucleic acid sequence of said vector result in production of a polyunsaturated fatty acid by said plant cell, plant or plant tissue.

10. The plant cell, plant or plant tissue of claim 9 wherein said polyunsaturated fatty acid is selected from the group consisting of ω

6-docosapentaenoic acid and docosahexaenoic acid.

11. A transgenic plant comprising said vector of claim 7, wherein expresion of said nucleic acid sequence of said vector results in production of a polyunsaturated fatty acid in seeds of said transgenic plant.