Strains for the production of flavonoids from glucose
The invention relates to the production of flavonoids and flavonoid precursors in cells through recombinant expression of tyrosine ammonia lyase (TAL), 4-coumarate:CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI).
1. An isolated cell that recombinantly expresses genes encoding tyrosine ammonia lyase (TAL), 4-coumarate:CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI), and wherein the cell produces one or more flavonoids or naringenin from glucose without additional precursor supplementation, and wherein the expression of the genes encoding TAL, 4CL, CHS and CHI is balanced in the cell by constitutive expression of CHS and CHI.
2. The isolated cell of claim 1 , wherein the gene encoding TAL is a yeast gene or a bacterial gene.
3. The isolated cell of claim 1 , wherein the gene encoding 4CL is a plant gene or a bacterial gene.
4. The isolated cell of claim 1 , wherein the at least one of the gene encoding CHS and the gene encoding CHI is a plant gene.
5. The isolated cell of claim 1 , wherein the genes encoding at least one of TAL, 4CL, CHS, and CHI are expressed from a single polycistronic operon, or wherein each of the genes is expressed from a separate promoter.
6. The isolated cell of claim 1 , wherein the cell is a prokaryotic cell.
7. The isolated cell of claim 6 , wherein the endogenous L-tyrosine production is at least 250 milligrams/liter.
8. The isolated cell of claim 6 , wherein the cell is a bacterial cell.
9. The isolated cell of claim 1 , wherein at least one of the genes encoding TAL, 4CL, CHS, and CHI is a synthetic gene that is codon optimized for expression in bacteria.
10. The isolated cell of claim 1 , wherein the cell is a eukaryotic cell.
11. The isolated cell of claim 1 , wherein at least one of the genes encoding TAL, 4CL, CHS, and CHI are expressed on plasmids, or wherein at least one of the genes encoding TAL, 4CL, CHS, and CHI are integrated into the genome of the cell.
12. The isolated cell of claim 1 , wherein the production of naringenin is increased by protein engineering of at least one of the TAL, 4CL, CHS, and CHI in the cell.
13. The isolated cell of claim 1 , wherein the cell further comprises a recombinantly-expressed malonate assimilation pathway.
14. The isolated cell of claim 1 , wherein the cell further comprises simultaneous deletions of genes encoding succinate dehydrogenase, alcohol/acetaldehyde dehydrogenase, branched chain amino acid transporter, and citrate lyase and overexpresses the enzymes acetyl-CoA synthase, acetyl-CoA carboxylase, biotin ligase, and pantothenate kinase.
15. The isolated cell of claim 1 , wherein upon culturing the cell produces at least 500 micrograms/liter naringenin in the culture medium.
16. A method for producing one or more flavonoids or naringenin comprising
culturing the cell of claim 1 to produce the one or more flavonoids or the naringenin.
17. The method of claim 16 , wherein the culture medium is not supplemented with a precursor of naringenin synthesis.
18. The method of claim 16 , wherein the cells are cultured in the presence of the fatty acid pathway inhibitor cerulenin or wherein the cell culture is supplemented with malonate.
19. The method of claim 16 , wherein the cells produce at least 500 micrograms/liter naringenin in the culture medium.