Yeasts modified to use carbon dioxide

Patent metadata

Field	Value
Publication number	US-10066234-B2
Application number	US-201515111410-A
Country	US
Kind code	B2
Filing date	Jan 16, 2015
Priority date	Jan 16, 2014
Publication date	Sep 4, 2018
Grant date	Sep 4, 2018

Abstract

The invention relates to yeast cells modified to express a functional type I RuBisCO enzyme, and a class II phosphoribulokinase. The expression of these enzymes recreates a Calvin cycle in said yeasts in order to enable the yeasts to use carbon dioxide.

First claim

The invention claimed is:

1. A transformed yeast cell, characterized in that it contains:
a) an expression cassette containing a sequence encoding the RbcL subunit of a bacterial form I RuBisCO enzyme, under the transcriptional control of a suitable promoter;
b) an expression cassette containing a sequence encoding the RbcS subunit of said RuBisCO enzyme, under the transcriptional control of a suitable promoter;
c) an expression cassette containing a sequence encoding the specific chaperone RbcX of said RuBisCO enzyme, under the transcriptional control of a suitable promoter;
d) an expression cassette containing a sequence encoding a general bacterial chaperone GroES, under the transcriptional control of a suitable promoter; and
e) an expression cassette containing a sequence encoding a general bacterial chaperone GroEL, under the transcriptional control of a suitable promoter, wherein the RbcX originates from a different bacteria than the GroES and the GroEL.

2. The transformed yeast cell according to claim 1 , characterized in that the GroES and GroEL originate-from E. coli.

3. The transformed yeast cell according to claim 1 , characterized in that the chaperone RbcX is a cyanobacterial chaperone.

4. The transformed yeast cell according to claim 1 , characterized in that GroES and GroEL come from a bacterium that does not naturally express a RuBisCO complex.

5. The transformed yeast cell according to claim 1 , characterized in that the three expression cassettes mentioned in points c), d), and e) of claim 1 form a continuous block of genetic information.

6. The transformed yeast cell according to claim 1 , characterized in that the expression cassettes mentioned in points c), d), and e) of claim 1 are carried by a single episomal genetic element.

7. The transformed yeast cell according to claim 1 , characterized in that said yeast is Saccharomyces cerevisiae.

8. The transformed yeast cell according to claim 1 , characterized in that the bacterial form I RuBisCO enzyme is a cyanobacterial RuBisCO enzyme.

9. The transformed yeast cell according to claim 8 , characterized in that said cyanobacterium belongs to the genus Synechococcus.

10. The transformed yeast cell according to claim 1 , characterized in that it further contains an expression cassette containing a sequence encoding a phosphoribulokinase (PRK) under the transcriptional control of a suitable promoter.

11. The transformed yeast cell according to claim 10 , characterized in that said PRK is a class II PRK.

12. The transformed yeast cell according to claim 11 , characterized in that said class II PRK is selected from PRKs from Spinacia oleracea, Euglena gracilis , or Synechococcus elongatus.

13. The transformed yeast cell according to claim 10 , characterized in that the promoter controlling transcription of the sequence encoding the PRK is an inducible promoter.

14. The transformed yeast cell according to claim 2 , characterized in that the chaperone RbcX is a cyanobacterial chaperone.

15. The transformed yeast cell according to claim 3 , characterized in that the GroES and GroEL come from a bacterium that does not naturally express a RuBisCO complex.

16. The transformed yeast cell according to claim 2 , characterized in that the three expression cassettes mentioned in points c), d), and e) of claim 1 form a continuous block of genetic information.

17. The transformed yeast cell according to claim 3 , characterized in that the three expression cassettes mentioned in points c), d), and e) of claim 1 form a continuous block of genetic information.

18. The transformed yeast cell according to claim 4 , characterized in that the three expression cassettes mentioned in points c), d), and e) of claim 1 form a continuous block of genetic information.

19. The transformed yeast cell according to claim 2 , characterized in that the expression cassettes mentioned in points c), d), and e) of claim 1 are carried by a single episomal genetic element.

20. The transformed yeast cell according to claim 2 , characterized in that the chaperone RbcX is a cyanobacterial chaperone.

21. The transformed yeast cell according to claim 20 , characterized in that the GroES and GroEL come from a bacterium that does not naturally express a RuBisCO complex.

22. The transformed yeast cell according to claim 20 , characterized in that the three expression cassettes mentioned in points c), d), and e) of claim 1 form a continuous block of genetic information.

23. The transformed yeast cell according to claim 1 , further containing:
f) an expression cassette containing a sequence encoding a class II PRK under the transcriptional control of a suitable promoter,

wherein the bacterial form I RuBisCO enzyme is a cyanobacterial RuBisCO enzyme and the GroES and GroEL are from E. coli.

Assignees

• Agronomique Inst Nat Rech
• Institut Nat Des Sciences Appliquees De Toulouse
• Centre Nat Rech Scient

Inventors

• Pompon Denis
• Paques Frederic
• Lesage Julie
• Guillouet Stephane
• Bonnot Florence
• Marc Jillian
• Gorret Nathalie
• Bideaux Carine
• Boutonnet Christel

Classifications

• C12N9/1205 Primary

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12Y207/01019

  Phosphoribulokinase (2.7.1.19) · CPC title

• C07K14/195

  from bacteria · CPC title

• C12N2830/002

  inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor · CPC title