Molecules associated with fatty acid biosynthetic pathways and uses thereof

Patent metadata

Field	Value
Publication number	US-10131897-B2
Application number	US-201514637356-A
Country	US
Kind code	B2
Filing date	Mar 3, 2015
Priority date	Mar 3, 2014
Publication date	Nov 20, 2018
Grant date	Nov 20, 2018

Abstract

The present disclosure relates in part to recombinant microorganisms that include non-native genes encoding PUFA-PKS polypeptides, and to methods of making and using such microorganisms for producing at least one PUFA. In particular, the disclosure further relates to methods and related materials useful for the production of at least one PUFA by heterologous expression of the nucleic acid sequences disclosed herein encoding PUFA-PKS polypeptides.

First claim

What is claimed is:

1. A recombinant eukaryotic microorganism comprising at least one non-native nucleic acid sequence encoding a prokaryotic Type A polyunsaturated fatty acid (PUFA) polyketide synthase (PKS) system comprising a pfaA gene, a pfaB gene, a pfaC gene, a pfaD gene, and a pfaE gene, wherein the pfaA gene encodes a polypeptide comprises comprising an amino acid sequence having at least 98% identity to SEQ ID NO:2,
the pfaB gene encodes a polypeptide comprises comprising an amino acid sequence having at least 96% identity to SEQ ID NO:4, the pfaC gene encodes a polypeptide comprises comprising an amino acid sequence having at least 98% identity to SEQ ID NO:6,
the pfaD gene encodes a polypeptide comprises comprising an amino acid sequence having at least 96% identity to SEQ ID NO:8, and
the pfaE gene encodes a polypeptide comprises comprising an amino acid sequence having at least 96% identity to SEQ ID NO: 10;
wherein the recombinant eukaryotic microorganism produces at least 50% more eicosapentaenoic acid (EPA) than a control eukaryotic microorganism.

2. A recombinant eukaryotic microorganism according to claim 1 , wherein the recombinant eukaryotic microorganism produces at least 75% more EPA than a control eukaryotic microorganism.

3. A recombinant eukaryotic microorganism according to claim 2 , wherein the eukaryotic microorganism produces at least 100% more EPA than a control eukaryotic microorganism.

4. A recombinant eukaryotic microorganism according to claim 1 , wherein the eukaryotic microorganism is a labyrinthulomycete.

5. A recombinant eukaryotic microorganism according to claim 4 , wherein the eukaryotic microorganism is of a genus selected from the group consisting of Aurantiochytrium, Oblongichytrium, Schizochytrium, Thraustochytrium , and Ulkenia.

6. A recombinant eukaryotic microorganism according to claim 1 , wherein the PUFA-PKS system is derived from a prokaryotic species of a genus selected from the group consisting of Shewanella, Vibrio, Aliivibrio, Pseudoaltermonas, Labyrinthuloides , and Psychromonas.

7. A recombinant eukaryotic microorganism according to claim 1 , wherein at least one non-native nucleic acid sequence is operably linked to a heterologous promoter.

8. A method for producing EPA, comprising culturing a recombinant eukaryotic microorganism according to claim 1 to produce EPA.

9. A method according to claim 8 , further comprising recovering EPA from the culture.

10. A method for according to claim 8 , wherein said culturing is performed at a temperature greater than 20° C.

11. A method for according to claim 10 , wherein said culturing is performed at a temperature greater than 25° C.

12. A method for according to claim 11 , wherein said culturing is performed at a temperature of at least 30° C.

Assignees

• Synthetic Genomics Inc
• Synthetics Genomics Inc

Inventors

• Toledo Gerardo V
• Xu Karen D
• Urano Jun

Classifications

• C12Y604/00

  Ligases forming carbon-carbon bonds (6.4) · CPC title

• C12P7/6427

  Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N9/93 Primary

  Ligases (6) · CPC title