Methods for decoupling yield and productivity of a non-catabolic compound produced by a host cell
Provided herein are compositions and methods for uncoupling the yield and productivity of an isoprenoid compound produced in a host cell. In some embodiments, the yield and productivity are uncoupled by genetically modifying the host cell to reduce flux through the citric acid cycle (TCA). In other embodiments, the yield and productivity are uncoupled by reducing the levels of ATP in the host cell.
1. A method of decoupling yield and productivity of an isoprenoid produced in a yeast cell capable of making the isoprenoid, wherein the method comprises: adding one or more ATP depleting agents to the yeast cell and reducing ATP levels during production of the isoprenoid.
2. The method of claim 1 , wherein the one or more ATP depleting agents is a weak organic acid.
3. The method of claim 2 , wherein the weak organic acid is selected from sorbic acid, acetic acid, benzoic acid, and propionic acid.
4. The method of claim 3 , wherein the weak organic acid is benzoic acid.
5. The method of claim 1 , wherein the ATP levels are reduced by over expression of one or more ATP dissipation enzymes.
6. The method of claim 5 , wherein the one or more ATP dissipation enzymes are selected from Saccharomyces cerevisiae SSB1 and ATP-diphosphohydrolase.
7. The method of claim 1 , wherein the ATP levels are reduced by over expression of one or more ATP uncoupling enzymes.
8. The method of claim 7 , wherein the one or more ATP uncoupling enzymes are selected from NADH oxidase (NOX) and alternative oxidase (AOX).
9. The method of claim 1 , wherein the ATP levels are reduced by expression of a futile cycle in the yeast cell.
10. The method of claim 9 , wherein the futile cycle is selected from simultaneous over expression of phosphofructokinase and fructose-1,6-bisphosphatase and simultaneous over expression of phosphoenolpyruvate carboxykinase and pyruvate carboxylase.
11. The method of claim 1 , wherein the isoprenoid is selected from the group consisting of a hemiterpene, monoterpene, diterpene, triterpene, tetraterpene, sesquiterpene, and polyterpene.
12. The method of claim 1 , wherein the isoprenoid is selected from the group consisting of abietadiene, amorphadiene, carene, α-farnesene, β-farnesene, farnesol, geraniol, geranylgeraniol, isoprene, linalool, limonene, myrcene, nerolidol, ocimene, patchoulol, β-pinene, sabinene, γ-terpinene, terpinolene, and valencene.
13. The method of claim 12 , wherein the isoprenoid is β-farnesene.
14. The method of claim 1 , wherein the yeast cell is a Saccharomyces cerevisiae.
15. The method of claim 1 , wherein the ATP levels are reduced by reducing carbon flux through the citric acid cycle (TCA) in the yeast cell.
16. The method of claim 15 , wherein carbon flux through the TCA cycle is reduced by decreasing expression or activity of one or more TCA enzymes.
17. The method of claim 16 , wherein the TCA enzymes are selected from citrate synthase, aconitate hydratase, NAD-dependent isocitrate dehydrogenase, 2-ketoglutarate dehydrogenase, succinyl-CoA ligase, succinate dehydrogenase, fumarate hydralase, peroxisomal malate dehydrogenase, and pyruvate carboxylase.