Delta-8 desaturase and its use in making polyunsaturated fatty acids

• US 7,256,033 B2
• Filed: 06/24/2005
• Issued: 08/14/2007
• Est. Priority Date: 06/25/2004
• Status: Expired due to Fees

• Current Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Damude, Howard Glenn, Zhu, Quinn Qun
• Primary Examiner(s): Prouty; Rebecca E.
• Assistant Examiner(s): CHOWDHURY, IQBAL HOSSAIN
• Application Number:
  US11/166,993
• Publication Number:
  US 20050287652A1
• Time in Patent Office: 781 Days
• Field of Search: 435/190, 435/134, 435/69.1, 435/320.1, 435/254.2, 435/483
• US Class Current: 435/252.3
• 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: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A23V 2300/21 Genetic modification
• 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 7/6432 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

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding a delta-8 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this delta-8 desaturase in plants and oleaginous yeast.

Claims

1. An isolated polynucleotide comprising:

(a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has the amino acid sequence as set forth in SEQ ID NO;

113;

or,

(b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.

2. The polynucleotide of claim 1 wherein the nucleotide sequence comprises SEQ ID NO:

112.

3. A recombinant construct comprising the polynucleotide of claim 1 operably linked to at least one regulatory sequence.

4. An isolated transformed cell comprising the isolated polynucleotide of either claim 1 or claim 2.

5. The cell of claim 4 wherein said cell is a yeast.

6. The cell of claim 5 wherein the yeast is an oleaginous yeast selected from the group of:

Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon and Lipomyces.

7. An oil-producing oleaginous yeast cell comprising a recombinant DNA construct comprising an isolated nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has the amino acid sequence as set forth in SEQ ID NO:

113.

8. The oil-producing oleaginous yeast of claim 7 wherein the yeast cell is an oleaginous yeast selected from the group of:

Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon and Lipomyces.

9. The oil-producing oleaginous yeast of claim 8 wherein the oleaginous yeast cell is a Yarrowia sp. and the oil comprises at least one polyunsaturated fatty acid selected from the group consisting of:

eicosatetraenoic acid and dihomo-γ

-linoleic acid.

10. The oil-producing oleaginous yeast of claim 7 wherein the recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide has the nucleotide sequence as set forth in SEQ ID NO:

112.