Diacylgylcerol acyl transferase (DGA1) polynucleotides, and methods of increasing yeast cell lipid production by overexpression of heterologous DGA1

• US 10,260,052 B2
• Filed: 02/24/2015
• Issued: 04/16/2019
• Est. Priority Date: 02/24/2014
• Status: Active Grant

• Current Assignee: Ginkgo Bioworks, Inc. (Ginkgo Bioworks Holdings, Inc.)
• Original Assignee: Novogy, Inc. (Ginkgo Bioworks Holdings, Inc.)
• Inventors: Brevnova, Elena E., Shaw, IV, Arthur J., Greenhagen, Emily H.
• Primary Examiner(s): Saidha, Tekchand
• Application Number:
  US15/120,952
• Publication Number:
  US 20160362663A1
• Time in Patent Office: 1,512 Days
• Field of Search: 435224, 435183, 435483
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6445 Glycerides
• CPC Class Codes: C12Y 203/0102 Diacylglycerol O-acyltransferase (2.3.1.20)
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

DGA1 catalyzes the final enzymatic step for converting acyl-CoA and 1,2-diacylglycerol to triacylglycerols (TAG) and CoA in yeast. Disclosed are methods for expression in an oleaginous yeast host of polynucleotide sequences encoding DGA1 from Rhodosporidium toruloides, Lipomyces starkeyi, Aurantiochytrium limacinium, Aspergillus terreus, or Claviceps purpurea. Also described herein are engineered recombinant host cells of Yarrowia lipolytica comprising heterologous DGA1 polynucleotides encoding DGA1 proteins, or functionally active portions thereof, having the capability of producing increased lipid production and possessing the characteristic of enhanced glucose consumption efficiency.

Claims

1. A method for producing a recombinant Yarrowia cell that has a higher lipid levels than a non-recombinant Yarrowia cell, the method comprising the steps of:

a) introducing into a Yarrowia cell a nucleic acid molecule encoding a heterologous polypeptide comprising SEQ ID NO;

9 or a biologically-active portion thereof; and

b) cultivating the recombinant Yarrowia cell, wherein the recombinant Yarrowia cell has a higher lipid content than a non-recombinant cell of the same species.

2. The method of claim 1, wherein the yeast cell is Y. lipolytica.

3. The method of claim 1, wherein the polynucleotide has at least 95% sequence identity to SEQ ID NO:

10.

4. The method of claim 1, wherein the polypeptide has at least 90% sequence identity to SEQ ID NO:

9.

5. The method of claim 1 wherein the biologically active portion is a 278 contiguous amino acids of SEQ ID NO:

9.