Genetically modified fungal cells and methods useful for producing prespatane

Patent metadata

Field	Value
Publication number	US-11845940-B2
Application number	US-202117187697-A
Country	US
Kind code	B2
Filing date	Feb 26, 2021
Priority date	Feb 27, 2020
Publication date	Dec 19, 2023
Grant date	Dec 19, 2023

Abstract

The present invention provides for a genetically modified fungal host cell capable of producing prespatane and/or epi-isozizaene comprising prespatane synthase (PPS) and/or epi-isozizaene synthase (EIZS).

First claim

What is claimed is:

1. A genetically modified fungal host cell capable of producing prespatane comprising prespatane synthase (PPS) wherein the PPS comprises an amino acid sequence having at least 70% identity with SEQ ID NO:1.

2. The genetically modified host cell of claim 1 , wherein the genetically modified host cell is a yeast.

3. The genetically modified host cell of claim 2 , wherein the genetically modified host cell is of the genus Rhodosporidium.

4. The genetically modified host cell of claim 3 , wherein the genetically modified host cell is Rhodosporidium toruloides.

5. The genetically modified host cell of claim 1 , wherein the PPS comprises an amino acid sequence having DDXXD (SEQ ID NO:3) or DDXXE (SEQ ID NO:4).

6. The genetically modified host cell of claim 5 , wherein the PPS comprises an amino acid sequence having at least 80% identity with SEQ ID NO:1.

7. The genetically modified host cell of claim 6 , wherein the PPS comprises an amino acid sequence having at least 90% identity with SEQ ID NO:1.

8. The genetically modified host cell of claim 7 , wherein the PPS comprises an amino acid sequence having at least 95% identity with SEQ ID NO:1.

9. The genetically modified host cell of claim 8 , wherein the PPS comprises an amino acid sequence having at least 99% identity with SEQ ID NO:1.

10. The genetically modified host cell of claim 9 , wherein the PPS comprises SEQ ID NO:1.

11. The genetically modified host cell of claim 1 , further comprising one or more enzymes of the mevalonate (MVA) pathway, wherein the MVA pathway is heterologous to the genetically modified host cell.

12. The genetically modified host cell of claim 11 , further comprising acetoacetyl-CoA thiolase (AtoB), HMG-CoA synthase (HMGS), HMG-CoA reductase (HMGR), mevalonate kinase (MK), phosphomevalonate kinase (PMK), mevalonate diphosphate decarboxylase (PMD), isopentenyl diphosphate (IPP) isomerase (Idi), and farnesyl diphosphate (FPP) synthase (IspA), which are heterologous to the genetically modified host cell.

13. A method for producing prespatane comprising: (a) providing a genetically modified host cell of claim 1 or a culture thereof, (b) culturing the genetically modified host cell to produce prespatane.

14. The method of claim 13 , wherein the culture comprises a biomass or hydrolysate thereof.

15. The method of claim 14 , wherein the biomass or hydrolysate thereof, is obtained from poplar.

16. The method of claim 13 , further comprising the step of extracting or separating the prespatane from the culture.

17. The method of claim 16 , further comprising the step of hydrogenating the prespatane extracted or separated from the culture.

18. The method of claim 16 , further comprising the step of introducing a fuel additive to the extracted or separated prespatane.

Assignees

• Univ California
• National Tech And Engineering Solutions Of Sandia Llc
• Nat Tech & Eng Solutions Sandia Llc

Inventors

• Geiselman Gina Marie
• Kirby James
• Lee Taek Soon
• Gladden John M
• Simmons Blake A

Classifications

• C12N15/815 Primary

  for yeasts other than Saccharomyces · CPC title

• C10L1/04

  essentially based on blends of hydrocarbons · CPC title

• C12N1/16

  Yeasts; Culture media therefor · CPC title

• C12N9/88 Primary

  Lyases (4.) · CPC title

• C12P5/007

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title