IP Library › Granted Patent US 12,398,398
Granted Patent B2
US 12,398,398 · App. 17/108,027 · Granted Aug 26, 2025

Recombinant yeast expressing rubisco and phosphoribulokinase

Inventors: Antonius Jeroen Adriaan Van Maris (Delft, NL); Jacobus Thomas Pronk (Delft, NL); Victor Gabriel Guadalupe Medina (Delft, NL); Hendrik Wouter Wisselink (Delft, NL)
Assignee: Danisco US Inc.
C12N15/81C07K14/245C12N1/16C12N9/1205C12N9/88C12P7/06C12P7/62C12P13/04C12Y207/01019C12Y401/01039Y02E50/10
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Quick Facts
Patent No.
US 12,398,398
App. No.
17/108,027
Granted
Aug 26, 2025
Kind
B2
Abstract

The invention relates to a recombinant yeast cell, in particular a transgenic yeast cell, functionally expressing one or more recombinant, in particular heterologous, nucleic acid sequences encoding ribulose-1,5-biphosphate carboxylase oxygenase (Rubisco) and phosphoribulokinase (PRK). The invention further relates to the use of carbon dioxide as an electron acceptor in a recombinant chemotrophic micro-organism, in particular a eukaryotic micro-organism.

Claims (35)

1. A recombinant yeast cell comprising:

one or more recombinant heterologous, nucleic acid sequences encoding Ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) and phosphoribulokinase (PRK), wherein the amino acid sequence of the Rubisco has at least 80% sequence identity with the sequence of SEQ ID NO: 2, and

the amino acid sequence of the PRK has at least 70% sequence identity with the sequence of SEO ID NO: 4; and

nucleic acid sequences encoding a GroEL

or a functional homologue of GroEL, and a GroES or a functional homologue of GroES, wherein

the respective GroEL and GroES homologues are functional as molecular chaperones,

wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 70% sequence identity with the sequence of SEO ID NO: 10; and

wherein the amino acid sequence of the GroES or the functional homologue of GroES has at least 70% sequence identity with the sequence of SEO ID NO: 12.

2. The recombinant yeast cell of claim 1 , wherein the Rubisco uses CO2 as an electron acceptor.

3. The recombinant yeast cell of claim 1 , wherein the Rubisco is a single subunit Rubisco.

4. The recombinant yeast cell of claim 1 , wherein the Rubisco is a prokaryotic form-II Rubisco.

5. The recombinant yeast cell of claim 1 , wherein the PRK is a PRK originating from a eukaryote.

6. The recombinant yeast cell of claim 5 , wherein the PRK originates from a Caryophyllales plant.

7. The recombinant yeast cell of claim 6 , wherein the Caryophyllales plant is Amaranthaceae or Spinacia.

8. The recombinant yeast cell of claim 1 , wherein the genus of said yeast cell is selected from the group consisting of Saccharomycesaceae, Schizosaccharomyces, Torulaspora, Kluyveromyces, Pichia, Zygosaccharomyces, Brettanomyces, Metschnikowia, Issatchenkia, Kloeckera , and Aureobasidium.

9. The recombinant yeast cell of claim 8 , wherein the genus of yeast cell is Saccharomycesceae.

10. The recombinant yeast cell of claim 9 , wherein the yeast cell is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces pastorianus, Saccharomyces beticus, Saccharomyces fermentati, Saccharomyces paradoxus, Saccharomyces uvarum , and Saccharomyces bayanus.

11. A method for preparing an alcohol under anaerobic conditions comprising fermenting a carbon source and the recombinant yeast cell of claim 1 .

12. The recombinant yeast cell of claim 7 , wherein the PRK is a Spinacia PRK.

13. The recombinant yeast cell of claim 9 , wherein the yeast cell is a Saccharomyces cerevisiae.

14. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 75% sequence identity with the sequence of SEQ ID NO: 10; and the amino acid sequence of the GroES or the functional homologue of GroES has at least 75% sequence identity with the sequence of SEQ ID NO: 12.

15. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 80% sequence identity with the sequence of SEQ ID NO: 10; and the amino acid sequence of the GroES or the functional homologue of GroES has at least 80% sequence identity with the sequence of SEQ ID NO: 12.

16. The recombinant yeast cell of claim 1 , further comprising a heterologous nucleic acid sequence encoding a dehydrogenase.

17. The recombinant yeast cell of claim 16 , wherein the dehydrogenase is a xylitol dehydrogenase.

18. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 85% sequence identity with the sequence of SEQ ID NO: 10; and the amino acid sequence of the GroES or the functional homologue of GroES has at least 85% sequence identity with the sequence of SEQ ID NO: 12.

19. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 90% sequence identity with the sequence of SEQ ID NO: 10; and the amino acid sequence of the GroES or the functional homologue of GroES has at least 90% sequence identity with the sequence of SEQ ID NO: 12.

20. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the GroEL or the functional homologue of GroEL has at least 95% sequence identity with the sequence of SEQ ID NO: 10; and the amino acid sequence of the GroES or the functional homologue of GroES has at least 95% sequence identity with the sequence of SEQ ID NO: 12.

21. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the Rubisco has at least 85% sequence identity with the sequence of SEQ ID NO: 2.

22. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the Rubisco has at least 90% sequence identity with the sequence of SEQ ID NO: 2.

23. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the Rubisco has at least 95% sequence identity with the sequence of SEQ ID NO: 2.

24. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the PRK has at least 75% sequence identity with the sequence of SEQ ID NO: 4.

25. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the PRK has at least 80% sequence identity with the sequence of SEQ ID NO: 4.

26. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the PRK has at least 85% sequence identity with the sequence of SEQ ID NO: 4.

27. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the PRK has at least 90% sequence identity with the sequence of SEQ ID NO: 4.

28. The recombinant yeast cell of claim 1 , wherein the amino acid sequence of the PRK has at least 95% sequence identity with the sequence of SEQ ID NO: 4.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 14, 2023
From: DSM IP ASSETS B.V.
To: DANISCO US INC.
Reel/Frame 066005/0848 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 1, 2020
From: VAN MARIS, ANTONIUS JEROEN ADRIAAN; PRONK, JACOBUS THOMAS; GUADALUPE MEDINA, VICTOR GABRIEL; WISSELINK, HENDRIK WOUTER
To: TECHNISCHE UNIVERSITEIT DELFT
Reel/Frame 054499/0488 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 1, 2020
From: TECHNISCHE UNIVERSITEIT DELFT
To: DSM IP ASSETS B.V.
Reel/Frame 054499/0695 →
Priority Claims (1)
EP 13156448 · Feb 22, 2013 · regional
Continuity (3)
Continuation 15980413 · May 15, 2018
Continuation 14767661
Related Publication 20210147859A1 · May 20, 2021
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