Modified microorganisms to increase yield of xylose-derived products

Patent metadata

Field	Value
Publication number	US-12584151-B1
Application number	US-202217880372-A
Country	US
Kind code	B1
Filing date	Aug 3, 2022
Priority date	Aug 3, 2022
Publication date	Mar 24, 2026
Grant date	Mar 24, 2026

Abstract

Methods and engineered hosts are disclosed that convert a lignocellulosic xylose-containing biomass source into xylonic acid and/or xylonate, which can be further processed into other useful derivatives. In particular, an exemplary engineered Bacidiomycetes, e.g., R. toruloides, host produces/expresses one or more fungal enzymes that convert xylose into xylonic acid/xylonate. Methods of using such hosts to consume pretreated lignocellulosic biomass in combination with certain native promoters and heterologous genes are also described herein.

First claim

It is claimed:

1 . An engineered organism comprising an engineered host derived from a Basidiomycete organism having a gene with at least 95% sequence identity to SEQ ID NO: 1 in its genome, the engineered host having the gene with 95% or greater sequence identity to SEQ ID NO: 1 deleted from its genome;
wherein XylB and XylC are expressed in the engineered host; and the engineered organism is capable of producing xylonic acid, xylonate, or a combination thereof.

2 . The engineered organism of claim 1 , wherein the engineered host is Rhodosporidium having the gene with 95% or greater sequence identity to SEQ ID NO: 1 deleted from its genome.

3 . The engineered organism of claim 2 , wherein the engineered host is R, toruloides with the gene with at least 99% sequence identity to SEQ ID NO: 1 deleted from its genome.

4 . The engineered organism of claim 1 , wherein XylB and XylC are codon optimized for the engineered host.

5 . A fermentation broth composition, comprising an energy source comprising xylose and an engineered host, the engineered host being derived from a Basidiomycete organism having a gene with at least 95% sequence identity to SEQ ID A NO: 1 in its genome, and the engineered host having the gene with at least 95% sequence identity to SEQ ID NO: 1 deleted from its genome;
wherein XylB and XylC are expressed in the engineered host; and the fermentation is capable of producing xylonate, or a combination thereof.

6 . The fermentation broth composition of claim 5 , wherein the engineered host is Rhodosporidium having the gene with at least 95% sequence identity to SEQ ID NO: 1 deleted from its genome.

7 . The fermentation broth composition of claim 6 , wherein the engineered host is R, toruloides having the gene with at least 95% sequence identity to SEQ ID NO: 1 deleted from its genome.

8 . The fermentation broth composition of claim 5 , wherein the fermentation broth composition has a pH of about 5.5 to about 6.

9 . The fermentation broth composition of claim 5 , wherein the energy source is a biomass hydrolysate.

10 . The fermentation broth composition of claim 5 , wherein gene sequences XylB and XylC are codon optimized for the engineered host.

11 . The fermentation broth composition of claim 5 , wherein the energy source is a corn stover hydrolysate.

12 . A method of making a xylose-derived product via a fermentation broth, comprising:
expressing XylB and XylC in the fermentation broth via introducing promoter sequences for expressing XylB and XylC in an engineered host: combining an energy source comprising xylose and the engineered host, the engineered host being derived from a Basidiomycete organism having a gene with at least A 95% sequence identity to SEQ ID NO: 1 in its genome, and the engineered host A having the gene with at least 95% sequence identity to SEQ ID NO: 1 deleted from its genome; and the method produces xylonic acid, xylonate, or a combination thereof.

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

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

15 . The method of claim 12 , wherein the engineered host is derived from Rhodosporidium.

16 . The method of claim 12 , wherein the engineered host is R, toruloides having SEQ ID NO: 1 deleted from its genome.

17 . The method of claim 12 , further comprising adding a nitrogen source to the fermentation broth.

18 . The method of claim 12 , wherein the fermentation broth has a pH of about 5.5 to about 6.

19 . The method of claim 12 , wherein the method yields 80% to 99% by weight of theoretical yield of xylonic acid, xylonate, or a combination thereof.

20 . The method of claim 12 , wherein XylB and XylC are codon optimized for the engineered host.

Assignees

• Nat Tech & Eng Solutions Sandia Llc

Inventors

• Adamczyk Paul Alexander
• Gladden John Michael
• Coradetti Samuel

Classifications

• C12N15/80

  for fungi · CPC title

• C12P2203/00

  Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source · CPC title

• C12P2201/00

  Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis · CPC title

• C12Y101/01307

  D-Xylose reductase (1.1.1.307) · CPC title

• C12N1/145

  Fungi isolates · CPC title