Mortierella alpina glycerol-3-phosphate o-acyltransferase for alteration of polyunsaturated fatty acids and oil content in oleaginous organisms

• US 7,192,762 B2
• Filed: 10/19/2005
• Issued: 03/20/2007
• Est. Priority Date: 11/04/2004
• 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: Xue, Zhixiong, Macool, Daniel Joseph
• Primary Examiner(s): Prouty; Rebecca E.
• Assistant Examiner(s): CHOWDHURY, IQBAL HOSSAIN
• Application Number:
  US11/254,173
• Publication Number:
  US 20060094091A1
• Time in Patent Office: 517 Days
• Field of Search: 435/252.3, 435/69.1, 435/320.1, 435/193, 536/23.2
• US Class Current: 435/252.3
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

Glycerol-3-phosphate o-acyltransferase (GPAT) participates in the first step of oil biosynthesis and is expected to play a key role in altering the quantity of long-chain polyunsaturated fatty acids (PUFAs) produced in oils of oleaginous organisms. The present application provides a nucleic acid fragment isolated from Mortierella alpina encoding a GPAT that is suitable for use in the manufacture of oils enriched in omega fatty acids in oleaginous organisms. Most desirably, the substrate specificity of the instant GPAT will be particularly useful to enable accumulation of long-chain PUFAs having chain lengths equal to or greater than C₂₀ in oleaginous yeast, such as Yarrowia lipolytica.

Claims

1. An isolated nucleic acid molecule selected from the group consisting of:

(a) an isolated nucleic acid molecule encoding a glycerol-3-phosphate o-acyltransferase, having the amino acid sequence as set forth in SEQ ID NO;

2;

(b) an isolated nucleic acid molecule encoding a glycerol-3-phosphate o-acyltransferase, that hybridizes with (a) under the following hybridization conditions;

0.1×

SSC, 0.1% SDS, 65°

C. and washed with 2×

SSC, 0.1% SDS followed by 0.1×

SSC, 0.1% SDS;

or,

an isolated nucleic acid molecule that is completely complementary to the full length of (a) or (b).

2. The isolated nucleic acid molecule of claim 1 as set forth in SEQ ID NO:

1.

4. A chimeric gene comprising the isolated nucleic acid molecule of claim 1 operably linked to suitable regulatory sequences.

5. A transformed host cell comprising the chimeric gene of claim 4, wherein the transformed host cell is selected from the group consisting of algae, bacteria, fungi and yeasts.

6. The transformed host cell of claim 5, wherein the yeast is an oleaginous yeast.

7. The transformed host cell of claim 6, wherein the oleaginous yeast cell is selected from the group consisting of:

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

8. The transformed host cell of claim 7, wherein to host cell is Yarrowia lipolytica.

3. An isolated nucleic acid molecule comprising a first nucleotide sequence encoding a glycerol-3-phosphate o-acyltransferase of at least 716 amino acids that has at least 95% identity based on the BLAST method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO:

2;

or a second nucleotide sequence comprising the complement of the first nucleotide sequence.