Novel acyltranserases and methods of using

Patent metadata

Field	Value
Publication number	US-2016237448-A1
Application number	US-201414576135-A
Country	US
Kind code	A1
Filing date	Dec 18, 2014
Priority date	Dec 18, 2013
Publication date	Aug 18, 2016
Grant date	—

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, comprising:
introducing a transgene into the organism, wherein the transgene comprises at least one nucleic acid sequence encoding an acyltransferase, wherein the at least one acyltransferase exhibits a substrate specificity for saturated fatty acids; thereby producing TAGs comprising MCFAs in the organism.

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 at least one acyltransferase is a lysophosphatidic acid acyltransferase (LPAT) or a diacylglycerol acyltransferase (DGAT).

5 . The method of claim 1 , wherein the at least one acyltransferase is a lysophosphatidic acid acyltransferase (LPAT) and a diacylglycerol acyltransferase (DGAT).

6 . The method of claim 4 , wherein the nucleic acid sequence encoding the LPAT is selected from the group consisting of a sequence having at least 95% sequence identity to SEQ ID NO:1 and a sequence having at least 95% sequence identity to SEQ ID NO:3.

7 . The method of claim 4 , wherein the nucleic acid sequence encoding the DGAT is selected from the group consisting of a sequence having at least 95% sequence identity to SEQ ID NO:7 and a sequence having at least 95% sequence identity to SEQ ID NO:9.

8 . The method of claim 1 , wherein the nucleic acid sequence encoding the at least one acyltransferase is selected from the group consisting of a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:1, a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:3, a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:7, and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:9.

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

10 . The method of claim 9 , 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.

11 . The method of claim 1 , wherein the organism is selected from the group consisting of a plant and a microbe.

12 . The method of claim 11 , wherein the plant is Camelina sativa.

13 . The method of claim 1 , wherein the transgene comprises a promoter.

14 . The method of claim 13 , wherein the promoter is a seed-specific promoter.

15 . The method of claim 11 , wherein the at least one nucleic acid sequence encoding an acyltransferase is operably linked to a seed-specific promoter.

16 . The method of claim 15 , wherein the medium-chain fatty acids are produced in the seed.

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

18 . A method of producing triacylglycerols (TAGs) comprising medium-chain fatty acids (MCFAs), comprising:
providing an organism comprising a transgene, wherein the transgene comprises at least one nucleic acid sequence encoding an acyltransferase, wherein the at least one acyltransferase exhibits a substrate specificity for saturated fatty acids; growing the organism under appropriate conditions; and obtaining TAGs comprising MCGAs from the organism.

19 . The method of claim 17 , wherein the TAGs are used in biofuel, jet fuel, detergents, and chemical feedstocks.

20 . A method of increasing the amount of triacylglycerols (TAGs) comprising medium-chain fatty acids (MCFAs) in the seed oil of a plant, comprising:
providing a plant comprising a nucleic acid encoding a FatB polypeptide; introducing a heterologous nucleic acid molecule into the plant comprising at least one nucleic acid sequence encoding an acyltransferase, wherein the at least one acyltransferase exhibits a substrate specificity for saturated fatty acids, thereby increasing the amount of TAGs comprising MCFAs in the seed oil of the plant without significantly changing the total oil content in the seed.

Assignees

• Univ Nebraska

Inventors

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

Classifications

• C12Y203/01051

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

• C12Y301/02

  Thioester hydrolases (3.1.2) · CPC title

• C07K14/415

  from plants · CPC title

• C12Y203/0102

  Diacylglycerol O-acyltransferase (2.3.1.20) · CPC title

• C12P7/6463

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