Production of isoprenoids
The present invention relates to a process for the production of isoprenoids Coenzyme Q-10 by microorganisms. More particularly, the present invention relates to a process for increased production of Coenzyme Q-10 by microorganisms of the genus Rhodobacter , preferably of the species R. sphaeroides which have been transformed with one or more gene(s) of the mevalonate (mev) operon from a different microorganism, preferably of the genus Paracoccus , more preferably of the species P. zeaxanthinifaciens , whereby the mev operon is mutated leading to an increased coenzyme Q-10 production. Sequences carrying such a mutation as well as a microorganism carrying such a mutated mev operon are also included.
1. An isolated polynucleotide comprising genes encoding a protein having hydroxymethylglutaryl-CoA reductase activity, isopentenyl diphosphate isomerase activity, hydroxymethylglutaryl-CoA synthase activity, mevalonate kinase activity, phosphomevalonate kinase activity, and diphosphomevalonate decarbo-xylase activity, wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of nucleotide sequences obtainable from SEQ ID NO:1 or 2 by mutating the cytosine at position 2949 to adenine and/or the thymidine at position 6948 to cytosine.
2. An isolated polynucleotide comprising SEQ ID NO:3 or a fragment thereof, wherein the polynucleotide encodes a protein having hydroxymethylglutaryl-CoA synthase activity.
3. The polynucleotide according to claim 2 comprising SEQ ID NO:3.
4. An isolated polynucleotide selected from the group consisting of:
(a) polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:7 and
(b) polynucleotides comprising the nucleotide sequence of SEQ ID NO:6.
5. A recombinant microorganism comprising a polynucleotide according to claim 1 .
6. A process for the production of isoprenoids comprising:
(a) introducing a polynucleotide according to claim 1 into a microorganism which is originally deficient in hydroxymethylglutaryl-CoA synthase activity, and
(b) cultivating the microorganism of step (a) under conditions to produce isoprenoids.
7. The process according to claim 6 wherein the microorganism in which the polynucleotide is introduced is selected from the group consisting of microorganisms belonging to the genus Rhodobacter.
8. The microorganism according to claim 5 further comprising a polynucleotide selected from the group consisting of:
(a) polynucleotides encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:9 and
(b) polynucleotides comprising the nucleotide sequence of SEQ ID NO:8.
9. The microorganism according to claim 5 which is selected from the group consisting of microorganisms belonging to the genus Rhodobacter.
10. A process of using a microorganism according to claim 5 , comprising cultivating the microorcianism under conditions to produce isoprenoids.
11. The process according to claim 6 wherein the isoprenoids comprise coenzyme Q10.
12. The process according to claim 7 wherein the microorganism belongs to the species R. sphaeroides.
13. The microorganism according to claim 9 wherein the microorganism belongs to the species R. sphaeroides.
14. The process according to claim 10 wherein the isoprenoids comprise coenzyme Q10.
15. The polynucleotide according to claim 1 which is comprised of SEQ ID NO:1 obtainable by mutating the cytosine at position 2949 to adenine and/or the thymidine at position 6948 to cytosine.
16. The polynucleotide according to claim 1 which is comprised of SEQ ID NO: 2 obtainable by mutating the cytosine at position 2949 to adenine and/or the thymidine at position 6948 to cytosine.
17. A process for the production of isoprenoids comprising:
(a) introducing a polynucleotide according to claim 2 into a microorganism which is originally deficient in hydroxymethylglutaryl-CoA synthase activity, and
(b) cultivating the microorganism of step (a) under conditions to produce isoprenoids.
18. A recombinant microorganism comprising a polynucleotide according to claim 2 .
19. A process of using a microorganism according to claim 18 , comprising cultivating the microorganism under conditions to produce isoprenoids.
20. A process for the production of isoprenoids comprising:
(a) introducing a polynucleotide according to claim 4 into a microorganism which is originally deficient in hydroxymethylglutaryl-CoA synthase activity, and
(b) cultivating the microorganism of step (a) under conditions to produce isoprenoids.
21. A recombinant microorganism comprising a polynucleotide according to claim 4 .
22. A process of using a microorganism according to claim 21 , comprising cultivating the microorganism under conditions to produce isoprenoids.
23. A process of using a microorganism according to claim 8 , comprising cultivating the microorganism under conditions to produce isoprenoids.