Methods and compositions for synthesis of long chain poly-unsaturated fatty acids

• US 6,410,288 B1
• Filed: 07/29/1999
• Issued: 06/25/2002
• Est. Priority Date: 04/11/1997
• Status: Expired due to Term

• Current Assignee: Abbott Laboratories Incorporated, Calgene Incorporated (Pharmacia Corp.)
• Original Assignee: Abbott Laboratories Incorporated, Calgene Incorporated (Pharmacia Corp.)
• Inventors: Mukerji, Pradip, Huang, Yung-Sheng, Thurmond, Jennifer, Chaudhary, Sunita, Knutzon, Deborah
• Primary Examiner(s): Nashed, Nashaat T.
• Application Number:
  US09/363,526
• Time in Patent Office: 1,062 Days
• Field of Search: 435/189, 435/254.2, 435/320.1, 435/254.21, 536/23.2
• US Class Current: 435/189
• 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 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A23L 33/40 Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
• 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 17/00 Drugs for dermatological disorders
• 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 7/6431 Linoleic acids [18:2[n-6]]
• 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 present invention relates to fatty acid desaturases able to catalyze the conversion of oleic acid to linoleic acid, linoleic acid to gamma-linolenic acid, or of alpha-linolenic acid to stearidonic acid. Nucleic acid sequences encoding desaturases, nucleic acid sequences which hybridize thereto, DNA constructs comprising a desaturase gene, and recombinant host microorganism or animal expressing increased levels of a desaturase are described. Methods for desaturating a fatty acid and for producing a desaturated fatty acid by expressing increased levels of a desaturase are disclosed. Fatty acids, and oils containing them, which have been desaturated by a desaturase produced by recombinant host microorganisms or animals are provided. Pharmaceutical compositions, infant formulas or dietary supplements containing fatty acids which have been desaturated by a desaturase produced by a recombinant host microorganism or animal also are described.

Claims

1. An isolated and purified Mortierella alpina polypeptide which removes hydrogen atoms from carbons 6 and 7 as numbered from the carboxy terminus of a fatty acid molecule to form a double bond.

2. The polypeptide of claim 1 wherein said polypeptide is isolated from a host cell wherein said host has been transformed with DNA encoding said polypeptide.

3. The polypeptide of claim 2 wherein said host cell is selected from the group consisting of a mammalian cell, a fungal cell, a yeast cell, an avian cell and an algal cell.

4. The polypeptide of claim 3 wherein said host cell is a yeast cell.

5. An isolated and purified Mortierella alpina polypeptide which removes hydrogen atoms from carbons 12 and 13 as numbered from the carboxy terminus of a fatty acid molecule to form a double bond.

6. The polypeptide of claim 5 wherein said polypeptide is isolated from a host cell wherein said host cell has been transformed with DNA encoding said polypeptide.

7. The polypeptide of claim 6 wherein said host cell is selected from the group consisting of a mammalian cell, a fungal cell, a yeast cell, an avian cell and an algal cell.

8. The polypeptide of claim 7 wherein said host cell is a yeast cell.

9. An isolated and purified Mortierella alpina polypeptide which removes hydrogen atoms from carbons 12 and 13 as numbered from the carboxy terminus of a fatty acid molecule to form a double bond, wherein the sequence of said polypeptide comprises a sequence selected from the group consisting of amino acid residues 50-53, 39-43, 172-176, 204-213 and 390-402 of SEQ ID NO:

2.

10. The polypeptide of claim 9 wherein said polypeptide is isolated from a host cell wherein said host cell has been transformed with DNA encoding said polypeptide.

11. The polypeptide of claim 10 wherein said host cell is selected from the group consisting of a mammalian cell, a fungal cell, a yeast cell, an avian cell and an algal cell.

12. The polypeptide of claim 11 wherein said host cell is a yeast cell.

13. An isolated and purified polypeptide encoded by a nucleotide sequence depicted in SEQ ID NO:

1 or SEQ ID NO;

3.

14. The polypeptide of claim 13 wherein said polypeptide is isolated from a host cell and wherein said host cell has been transformed with DNA compromising said nucleotide sequence.

15. The polypeptide of claim 14 wherein said host cell is selected from the group consisting of a mammalian cell, a fungal cell, a yeast cell, an avian cell and an algal cell.

16. The polypeptide of claim 15 wherein said host cell is a yeast cell.

17. An isolated and purified polypeptide comprising an amino acid sequence depicted in SEQ ID NO:

2 or SEQ ID NO;

4.

18. The polypeptide of claim 17 wherein said polypeptide is isolated from a host cell and wherein said host has been transformed with DNA encoding said polypeptide.

19. The polypeptide of claim 18 wherein said host cell is selected from the group consisting of a mammalian cell, a fungal cell, a yeast cell, an avian cell and an algal cell.

20. The polypeptide of claim 19 wherein said host cell is a yeast cell.