Acyltransferases and methods of using

Patent metadata

Field	Value
Publication number	US-10280431-B2
Application number	US-201414576135-A
Country	US
Kind code	B2
Filing date	Dec 18, 2014
Priority date	Dec 18, 2013
Publication date	May 7, 2019
Grant date	May 7, 2019

Abstract

Provided herein are novel acyltransferases and methods of using such novel acyltransferases in making medium-chain fatty acids.

First claim

What is claimed is:

1. A method of producing triacylglycerols (TAGs) comprising medium-chain fatty acids (MCFAs) in an organism, said method comprising:
introducing a first transgene into the organism, wherein the first transgene comprises at least one nucleic acid sequence encoding a lysophosphatidic acid acyltransferase (LPAT) that exhibits a substrate specificity for saturated fatty acids, wherein the nucleic acid sequence encoding the LPAT has at least 95% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 3; and
introducing a second transgene into the organism, wherein the second transgene comprises at least one nucleic acid sequence encoding a diacylglycerol acyltransferase (DGAT) that exhibits a substrate specificity for saturated fatty acids, wherein the nucleic acid sequence encoding the DGAT has at least 95% sequence identity to SEQ ID NO: 7 or SEQ ID NO: 9,
thereby producing TAGs comprising MCFAs in the organism, wherein the organism is a plant or a microbe.

2. The method of claim 1 , wherein at least 20% of the TAGs comprising MCFAs have a C8:0 or a C10:0 at sn-2 position.

3. The method of claim 1 , wherein the saturated fatty acids are selected from the group consisting of C8:0 and C10:0.

4. The method of claim 1 , wherein the nucleic acid sequence encoding the LPAT is a sequence having at least 95% sequence identity to SEQ ID NO:1.

5. The method of claim 1 wherein the nucleic acid sequence encoding the DGAT is a sequence having at least 95% sequence identity to SEQ ID NO:7.

6. The method of claim 1 , wherein the organism further comprises a nucleic acid sequence encoding a medium-chain fatty acid (MCFA)-specific thioesterase FatB.

7. The method of claim 6 , wherein the nucleic acid sequence encoding the MCFA-specific thioesterase FatB is selected from the group consisting of a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:11, a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:13, and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:15.

8. The method of claim 1 , wherein the organism is a plant, and wherein the plant is Camelina sativa.

9. The method of claim 1 , wherein the first transgene and the second transgene each comprises a promoter.

10. The method of claim 9 , wherein the organism is a plant, and wherein each promoter is a seed-specific promoter.

11. The method of claim 1 , wherein the at least one nucleic acid sequence encoding a LPAT is operably linked to a seed-specific promoter, and wherein the at least one nucleic acid sequence encoding a DGAT is operably linked to a seed-specific promoter.

12. The method of claim 11 , wherein the medium-chain fatty acids are produced in the seed.

13. The method of claim 1 , wherein the introducing step is performed using Agrobacterium transformation, particle bombardment, or electroporation of protoplasts.

14. A method of producing triacylglycerols (TAGs) comprising medium-chain fatty acids (MCFAs), comprising:
providing an organism comprising a first and a second transgene, wherein the organism is a plant or a microbe,
wherein the first transgene comprises at least one nucleic acid sequence encoding a lysophosphatidic acid acyltransferase (LPAT) that exhibits a substrate specificity for saturated fatty acids, wherein the nucleic acid sequence encoding the LPAT has at least 95% sequence identity to SEQ ID NO:1 or SEQ ID NO:3,
wherein the second transgene comprises at least one nucleic acid sequence encoding a diacylglycerol acyltransferase (DGAT) that exhibits a substrate specificity for saturated fatty acids, wherein the nucleic acid sequence encoding the DGAT has at least 95% sequence identity to SEQ ID NO:7 or SEQ ID NO:9;
growing the organism under appropriate conditions; and
obtaining TAGs comprising MCFAs from the organism.

15. The method of claim 14 , wherein the TAGs are used in biofuel, jet fuel, detergents, and chemical feedstocks.

Assignees

• Nutech Ventures

Inventors

• Cahoon Edgar
• Iskandarov Umidjon
• Kim Hae Jin
• Collins-Silva Jillian

Classifications

• C12Y203/0102

  Diacylglycerol O-acyltransferase (2.3.1.20) · CPC title

• C07K14/415

  from plants · CPC title

• C12Y301/02

  Thioester hydrolases (3.1.2) · CPC title

• C12Y203/01051

  1-Acylglycerol-3-phosphate O-acyltransferase (2.3.1.51) · CPC title

• C12P7/6463

  obtained from glyceride producing microorganisms, e.g. single cell oil · CPC title