Microbial engineering for the production of chemical and pharmaceutical products from the isoprenoid pathway

1. A method for increasing terpenoid production in a cell that produces one or more terpenoids, comprising

providing a bacterial cell that produces isopentenylpyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) through an upstream methylerythritol phosphate pathway (MEP) and converts the IPP and DMAPP to a terpenoid through a recombinantly expressed downstream terpenoid synthesis pathway; and

culturing the bacterial cell to produce the terpenoid, wherein accumulation of indole in the culture is controlled to below 100 mg/L to thereby increase terpenoid production.

2. The method of claim 1 , wherein accumulation of indole in the culture is controlled by balancing the upstream MEP pathway with the downstream terpenoid synthesis pathway.

3. The method of claim 2 , wherein the step of culturing comprises removing the accumulated indole.

4. The method of claim 1 , wherein the one or more terpenoids is a monoterpenoid, a sesquiterpenoid, a diterpenoid, a triterpenoid or a tetraterpenoid.

5. The method of claim 4 , wherein the one or more terpenoids is taxadiene or a taxol precursor.

6. The method of claim 2 , further comprising measuring the amount or concentration of indole in the culture.

7. The method of claim 6 , wherein the method comprises measuring the amount or concentration of indole two or more times.

8. The method of claim 6 , wherein the measured amount or concentration of indole is used to guide a process of producing one or more terpenoids.

9. The method of claim 6 , wherein the measured amount or concentration of indole is used to guide strain construction.

10. The method of claim 1 , wherein the method amplifies metabolic flux through IPP and DMAPP, and at least one intermediate selected from geranyl diphosphate (GPP), geranylgeranyl diphosphate (GGPP), farnesyl diphosphate (FPP), and farnesyl geranyl diphosphate (FGPP).

11. The method of claim 1 , wherein the bacterial cell recombinantly expresses a geranylgeranyl diphosphate synthase (GGPPS).

12. The method of claim 1 , wherein the bacterial cell has an amplified copy number of the dxs, idi, ispD and ispF genes of the MEP pathway.

13. The method of claim 1 , wherein the terpenoid is geraniol, citronellol, nootkatone, cineol, limonene, eleutherobin, sarcodictyin, a pseudopterosin, a ginkgolide, a stevioside, Rebaudioside A, sclareol, labdenediol, levopimaradiene, sandracopimaradiene or isopemaradiene.

14. The method of claim 1 , wherein the bacterial cell is Escherichia spp., Streptomyces spp., Zymomonas spp., Acetobacter spp., Citrobacter spp., Synechocystis spp., Rhizobium spp., Clostridium spp., Corynebacterium spp., Streptococcus spp., Xanthomonas spp., Lactobacillus spp., Lactococcus spp., Bacillus spp., Alcaligenes spp., Pseudomonas spp., Aeromonas spp., Azotobacter spp., Comamonas spp., Mycobacterium spp., Rhodococcus spp., Gluconobacter spp., Ralstonia spp., Acidithiobacillus spp., Microlunatus spp., Geobacter spp., Geobacillus spp., Arthrobacter spp., Flavobacterium spp., Serratia spp., Saccharopolyspora spp., Thermus spp., Stenotrophomonas spp., Chromobacterium spp., Sinorhizobium spp., Saccharopolyspora spp., Agrobacterium spp., or Pantoea spp.

15. The method of claim 1 , wherein the bacterial cell is Bacillus spp., Zymomonas spp., Corynebacterium spp., or Pseudomonas spp.

16. A method for producing a terpenoid, comprising

culturing a bacterial cell that produces isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) through an upstream methylerythritol phosphate pathway (MEP) and which converts the IPP and DMAPP to a terpenoid through a recombinantly expressed downstream terpenoid synthesis pathway, wherein the bacterial cell produces the terpenoid without accumulating indole in the culture media at greater than 100 mg/L;

wherein the bacterial cell has an amplified copy number of one or more of dxs, idi, ispD, and ispF, and wherein the terpenoid is selected from taxadiene or taxol precursor, citronellol, geraniol, nootkatone, cineol, limonene, eleutherobin, sarcodictyin, a pseudopterosin, a ginkgolide, a stevioside, Rebaudioside A, sclareol, labdenediol, levopimaradiene, sandracopimaradiene and isopemaradiene; and

recovering the terpenoid.