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1. WO2008008256 - PROCÉDÉS PERMETTANT D'AMÉLIORER LA PRODUCTION DE COMPOSÉS ISOPRÉNOÏDES PAR DES CELLULES HÔTES

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS

What is claimed is:

1. A method for producing an isoprenoid or isoprenoid precursor compound in a genetically modified host cell, the method comprising culturing the genetically modified host cell in a suitable medium, wherein the genetically modified host cell comprises a biosynthetic pathway that converts a substrate into isopentenyl pyrophosphate (IPP), wherein the genetically modified host cell comprises at least one synthetic intergenic region (IGR), wherein the at least one synthetic IGR is disposed between a set of two coding regions encoding two enzymes in the biosynthetic pathway.

2. The method of claim 1 , wherein the biosynthetic pathway is a 1 -deoxy-D-xylulose 5-diphosphate pathway that converts pyruvate and D-glyceraldehyde-3 -phosphate into IPP.

3. The method of claim 1, wherein the biosynthetic pathway is a mevalonate pathway that converts acetyl-CoA into IPP.

4. The method of claim 1 , wherein the synthetic IGR comprises a nucleotide sequence that forms a hairpin.

5. The method of claim 4, wherein the hairpin is a short hairpin.

6. The method of claim 4, wherein the hairpin is a long hairpin.

7. The method of claim 4, wherein the hairpin is part of a secondary structure selected from a stem-loop structure, a bulge-loop structure, a pseudoknot, a multi-loop structure, and an interior loop structure.

8. The method of claim 1 , wherein the synthetic IGR has a length of from about 15 nucleotides to about SOO nucleotides.

9. The method of claim 1 , wherein the synthetic IGR comprises a riboendonuclease recognition site.

10. The method of claim 9, wherein the riboendonuclease recognition site is an RNAse III recognition site.

11. The method of claim 3, wherein the riboendonuclease recognition site is an RNAse E recognition site.

12. The method of claim 1 , wherein the synthetic IGR comprises a nucleotide sequence having at least about 75% nucleotide sequence identity to the nucleotide sequence set forth in any one of SEQ ID NOs:7, 8, 11, 12, 15, 16, 19, 20, and 62-77.

13. The method of claim 3, wherein the biosynthetic pathway comprises one or more enzymes heterologous to the host cell.

14. The method of claim 3, wherein the two coding regions comprise a nucleotide sequence encoding acetoacetyl-CoA thiolase, and a nucleotide sequence encoding
hydroxymethylglutaryl-CoenzymeA synthase (HMGS).

15. The method of claim 3, wherein the two coding regions comprise a nucleotide sequence encoding hydroxymethylglutaryl-CoenzymeA synthase (HMGS), and a nucleotide sequence encoding hydroxymethylglutaryl-CoenzymeA reductase (HMGR).

16. The method of claim 1, wherein at least a second synthetic IGR is disposed between a second set of two coding regions in the biosynthetic pathway, wherein at least one coding region of the second set is different from a coding region of the first set.

17. The method of claim 1 , wherein the host cell is a prokaryotic cell .

18. The method of claim 13, wherein the host cell has a functionally disabled DXP pathway.

19. The method of claim 1 , wherein the host cell is a eukaryotic cell.

20. The method of claim 15, wherein the host cell is a yeast cell.

21. . The method of claim 1, further comprising recovering the isoprenoid or isoprenoid precursor from the cell and/or the culture medium.

22. A method for producing an isoprenoid or isoprenoid precursor compound in a genetically modified host cell, the method comprising culturing the genetically modified host cell in a suitable medium, wherein the genetically modified host cell comprises a biosynthetic pathway that converts a substrate into isopentenyl pyrophosphate (DPP), wherein the genetically modified host cell comprises at least one synthetic intergenic region (IGR) that comprises a nucleotide sequence that forms a hairpin, wherein the at least one synthetic IGR is disposed between a set of two coding regions encoding two enzymes in the biosynthetic pathway.

23. The method of claim 22, wherein the synthetic IGR comprises a nucleotide sequence having at least about 75% nucleotide sequence identity to the nucleotide sequence set forth in one of SEQ ID NOs:7, 8, 11, 12, 15, 16, 19, and 20.