Microbial fermentation for the production of terpenes
The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.
1. A recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell wherein the host cell is a Clostridium bacterium or a Moorella bacterium, wherein the nucleic acid is expressed in the host cell, and the recombinant cell is capable of producing isoprene from at least one Cl compound selected from the group consisting of carbon monoxide and carbon dioxide, wherein a nucleic acid encoding a group of exogenous enzymes acting in a mevalonate pathway is further introduced so that an isopentenyl diphosphate synthesis ability by a mevalonate pathway is further imparted, and wherein the group of exogenous enzymes acting in a mevalonate pathway comprises mevalonate kinase, mevalonate diphosphate decarboxylase, phosphomevalonate kinase, isopentenyl diphosphate (IPP) isomerase, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, and HMG-CoA synthase.
2. The recombinant cell according to claim 1 , having carbon monoxide dehydrogenase.
3. The recombinant cell according to claim 1 , wherein the mevalonate pathway is that of yeast or bacterium.
4. The recombinant cell according to claim 2 , wherein a nucleic acid encoding at least one enzyme acting in a DXS pathway is further introduced, and the nucleic acid is expressed in the host cell.
5. The recombinant cell according to claim 4 , wherein the DXS pathway is that of a different organism than the host cell.
6. The recombinant cell according to claim 1 , wherein the isoprene synthase is derived from a plant.
7. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase comprises SEQ ID NO: 21.
8. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase introduced into the host cell is codon optimized.
9. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase introduced into the host cell is incorporated in a genome of the host cell.
10. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase introduced into the host cell is incorporated in a plasmid.
11. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase and the nucleic acid encoding a group of exogenous enzymes acting in a mevalonate pathway are regulated by a constitutive promoter.
12. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isopentenyl diphosphate synthesis ability by a mevalonate pathway introduced into the host cell is codon optimized.
13. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isopentenyl diphosphate synthesis ability by a mevalonate pathway introduced into the host cell is incorporated in a genome of the host cell.
14. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isopentenyl diphosphate synthesis ability by a mevalonate pathway introduced into the host cell is incorporated in a plasmid.
15. A method for producing isoprene by culturing the recombinant cell according to claim 1 using at least one Cl compound selected from the group consisting of carbon monoxide and carbon dioxide as a carbon source, to allow the recombinant cell to produce isoprene.
16. The method according to claim 15 , wherein the recombinant cell is provided with a gas comprising hydrogen.
17. The method according to claim 15 , wherein the isoprene is recovered.
18. The method of claim 15 , wherein the Cl compound is derived from an industrial process selected from the group consisting of ferrous metal products manufacturing, non-ferrous products manufacturing, petroleum refining, coal gasification, electric power production, carbon black production, ammonia production, methanol production, and coke manufacturing.
19. The method of claim 15 , wherein the Cl compound is syngas.
20. A method for producing isoprene by bringing at least one Cl compound selected from the group consisting of carbon monoxide and carbon dioxide into contact with the recombinant cell according to claim 1 , to allow the recombinant cell to produce isoprene from the Cl compound.
21. The method according to claim 20 , wherein the recombinant cell is provided with a gas comprising hydrogen.
22. The method according to claim 20 , wherein the isoprene is recovered.