IP Library › Granted Patent US 12,215,373
Granted Patent B1
US 12,215,373 · App. 18/121,697 · Granted Feb 4, 2025

Modified yeast microorganisms to increase yield of 3-hydropropionic acid

Inventors: Di Liu (Emeryville, CA); Peter Britton Otoupal (Oakland, CA); HeeJin Hwang (Emeryville, CA); John Michael Gladden (Martinez, CA)
Assignee: National Technology & Engineering Solutions of Sandia, LLC
C12P7/52C12N1/205C12N9/22C12N2310/20
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Quick Facts
Patent No.
US 12,215,373
App. No.
18/121,697
Granted
Feb 4, 2025
Kind
B1
Abstract

Methods and compositions (e.g., engineered hosts) are disclosed for use in converting biomass to 3-hydropropionic acid. In particular embodiments, the methods include use of an engineered Rhodosporidium yeast, such as R. toruloides , the engineered R. toruloides having the RT04_8975 gene deleted from its genome, combined with a lignocellulosic hydrolysate, sourced, for example, from a biomass. A promoter for enhancing transport of 3HP is also incorporated by addition to the R. toruloides genome, for example, by modified lithium acetate transformation.

Claims (17)

1. An engineered Basidiomycete organism comprising a deletion of a gene comprising at least 90% sequence identity to the nucleotide sequence of SEQ ID NO: 1 in its genome, wherein the Basidiomycete organism is selected from the group consisting of: Rhodosporidium toruloides, Rhodosporidium azoricum, Rhodosporidium fluviale, Rhodosporidium lusitaniae, Rhodosporidium babievae, Rhodosporidium diobovatum, Rhodosporidium paludigenum, Rhodosporidium sphaerocarpum , and Rhodosporidium kratochvilovae.

2. The engineered Basidiomycete organism of claim 1 , wherein the engineered Basidiomycete organism is R. toruloides.

3. The engineered Basidiomycete organism of claim 1 , wherein the engineered Basidiomycete organism further comprises a transporter comprising the amino acid sequence of SEQ ID NO: 4.

4. The engineered Basidiomycete organism of claim 1 , wherein the engineered Basidiomycete organism expresses an Acetyl CoA carboxylase.

5. A fermentation broth composition comprising an energy source comprising glucose and the engineered Basidiomycete organism of claim 1 .

6. The fermentation broth composition of claim 5 , wherein the engineered Basidiomycete organism is R. toruloides.

7. The fermentation broth of claim 5 , wherein the engineered Basidiomycete organism further comprises a transporter comprising the amino acid sequence of SEQ ID NO: 4.

8. The fermentation broth of claim 5 , wherein the engineered hest Basidiomycete organism expresses an Acetyl CoA carboxylase.

9. The fermentation broth of claim 5 , further comprising a urea nitrogen source.

10. The fermentation broth of claim 5 , wherein the energy source further comprises a biomass hydrolysate.

11. The fermentation broth of claim 9 , wherein a C:N ratio in the fermentation broth is 4:1 to 8:1.

12. A method of producing 3 hydroxypropionic acid, comprising: combining an energy and material source and the engineered Basidiomycete organism of claim 1 , thereby producing 3 hydroxypropionic acid.

13. The method of claim 12 , further comprising introducing promoter sequences into the Basidiomycete organism to drive monocarboxylate transporter expression for promoting 3HP transport in the engineered host.

14. The method of claim 12 , wherein the energy source is a biomass hydrolysate.

15. The method of claim 12 , wherein the energy and material source is deacetylated mechanically refined corn stover.

16. The method of claim 12 , further comprising overexpressing a transporter comprising the amino acid sequence of SEQ ID NO: 4 in the engineered Basidiomycete organism hest.

17. The method of claim 16 , wherein the transporter is expressed under control of pGAPDH promoter, pTEF1 promoter, or both pGAPDH and pTEF1 promoters.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 20, 2025
From: POMRANING, KYLE R.; KIM, JOONHOON; MAGNUSON, JON K.; DAI, ZIYU
To: BATTELLE MEMORIAL INSTITUTE
Reel/Frame 070279/0857 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 16, 2023
From: LIU, DI; OTOUPAL, PETER BRITTON; HWANG, HEEJIN; GLADDEN, JOHN MICHAEL
To: NATIONAL TECHNOLOGY & ENGINEERING SOLUTIONS OF SANDIA, LLC
Reel/Frame 063971/0166 →
CONFIRMATORY LICENSE Recorded May 8, 2023
From: NATIONAL TECHNOLOGY & ENGINEERING SOLUTIONS OF SANDIA, LLC
To: U.S. DEPARTMENT OF ENERGY
Reel/Frame 063561/0141 →
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