Enzyme directed oil biosynthesis in microalgae

• US 8,362,318 B2
• Filed: 12/16/2009
• Issued: 01/29/2013
• Est. Priority Date: 12/18/2008
• Status: Active Grant

• Current Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Original Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Inventors: Benning, Christoph, Miller, Rachel, Moellering, Eric R.
• Primary Examiner(s): O Hara, Eileen B
• Application Number:
  US12/639,304
• Publication Number:
  US 20100192258A1
• Time in Patent Office: 1,140 Days
• Field of Search: None
• US Class Current: 800/278
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6409 Fatty acids
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12Y 203/0102 Diacylglycerol O-acyltransferase (2.3.1.20)

Abstract

The present invention is related to biosynthetic oil compositions and methods of making thereof. In some embodiments, the invention relates to the use of endogenous enzymes in plants capable of synthesizing oil. In preferred embodiments, said plants are algae. In further embodiments, said algae are from the family Chlamydomonas, Nannochloropsis, Dunaliella, Chlorella and Scenedesmus. In still further embodiments, said endogenous enzymes are diacylglycerol acyltransferases.

Claims

1. A method comprising:

a) providing;

i) algae, and

ii) a vector comprising a nucleic acid sequence encoding a diacylglycerol acyltransferase operably linked to a promoter, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

8 and SEQ ID NO;

11; and

b) transfecting said algae with said vector under conditions such that said diacylglycerol acyltransferase is expressed.

2. The method of claim 1, wherein said expression is under conditions such that triacylglycerol (TAG), free fatty acid and/or diacylglycerol (DAG) is produced.

3. The method of claim 2, further comprising c) recovering said TAG, said free fatty acid, and/or said DAG from said algae and/or oil from said algae.

4. The method of claim 1, wherein said expression is ectopic.

5. The method of claim 1, wherein said expression of said nucleic acid is upregulated between 1.5-3 fold relative to wild type expression of said nucleic acid.

6. The method of claim 1, wherein said algae is selected from the group consisting of Chlamydomonas, Nannochloropsis, Dunaliella, Chlorella and Scenedesmus.

7. The method of claim 1, wherein said algae is Chlamydomonas reinhardtii.

9. The algae of claim 6, wherein said promoter is a heterologous promoter.

10. The algae of claim 6, wherein said promoter is an inducible promoter.

8. Algae transfected with a nucleic acid encoding a diacylglycerol acyltransferase operably linked to a promoter, wherein said nucleic acid is selected from the group consisting of SEQ ID NO:

5, SEQ ID NO;

8 and SEQ ID NO;

11.

11. A non-algae host cell, wherein said cell comprises a nucleic acid sequence encoding an algae diacylglycerol acyltransferase operably linked to a promoter, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NO:

5, SEQ ID NO;

8 and SEQ ID NO;

11.

12. A method comprising:

a) providing;

i) a rutabaga, and

ii) a vector comprising a nucleic acid encoding a diacylglycerol acyltransferase operably linked to a promoter, wherein said nucleic acid is selected from the group consisting of SEQ ID NO;

5, SEQ ID NO;

8 and SEQ ID NO;

11;

b) transfecting said rutabaga with said vector under conditions such that said diacylglycerol acyltransferase is expressed.

13. A vector, comprising a nucleic acid encoding an algae diacylglycerol acyltransferase, wherein said nucleic acid is operably linked to a promoter, and wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NO:

5, SEQ ID NO;

8 and SEQ ID NO;

11.

14. The vector of claim 13, wherein said vector is a yeast shuttle vector.