Methods for recovering water-immiscible isoprenoid compounds from microbial biomass

1. A method for recovering one or more water immiscible compounds from a microbial biomass, comprising the steps of:

a. treating the microbial biomass by:

i. acidifying the microbial biomass at a pH of 2-4; and

ii. disrupting the microbial biomass;

to form an acidified, disrupted microbial biomass;

b. heating the acidified, disrupted microbial biomass to form a heated, acidified, disrupted microbial biomass;

c. contacting the heated, acidified, disrupted microbial biomass with a disulfonated surfactant in an amount sufficient to release at least 30% of the one or more water immiscible compounds from the microbial biomass;

d. recovering the one or more water immiscible compounds, wherein the water immiscible compounds comprise an isoprenoid, a polyketide, or a fatty acid.

2. The method of claim 1 , wherein the acidifying step precedes the disrupting step.

3. The method of claim 1 , wherein the disrupting step precedes the acidifying step.

4. The method of claim 1 , wherein the disrupting step and the acidifying step are simultaneous.

5. The method of claim 1 , wherein the acidifying step is at a pH of 2.5.

6. The method of claim 1 , wherein the disrupting step comprises mechanical disrupting, sonicating, freezing/thawing, grinding, chemical disrupting, enzymatic disrupting, or a combination thereof.

7. The method of claim 1 , wherein the disrupting step comprises homogenizing with one or more passes through a homogenizer at 600 bar to 1000 bar.

8. The method of claim 1 , wherein the disrupting step comprises homogenizing with one or more passes through a homogenizer at about 900 bar.

9. The method of any of claim 1 , wherein the heating step is at 60°-80° C.

10. The method of claim 1 , wherein the heating step is 70° C.

11. The method of claim 1 , wherein the disulfonated surfactant is a disulfonated phenyl ether detergent.

12. The method of claim 1 , wherein the disulfonated surfactant comprises:

or a salt thereof, wherein R is a C 6 -C 16 alkyl.

13. The method of claim 12 , wherein R is a C 12 alkyl.

14. The method of claim 1 , wherein the disulfonated surfactant is disodium lauryl phenyl ether disulfonate (DOWFAX 2A1™).

15. The method of claim 1 , wherein the heated, acidified, disrupted microbial biomass is contacted with 0.2 to 1.5% disulfonated surfactant.

16. The method of claim 1 , wherein the heated, acidified, disrupted microbial biomass is contacted with 1.0% disulfonated surfactant.

17. The method of claim 1 , wherein the recovering step comprises centrifugation, solvent extraction, chromatography, or a combination thereof.

18. The method of claim 1 , wherein at least 40, 50, 60, 70, or 75% of the one or more water immiscible compounds from the microbial biomass is released.

19. The method of claim 1 , comprising the steps of:

a. treating the microbial biomass by:

i. acidifying the microbial biomass to a pH of 2.5; and then

ii. disrupting the microbial biomass in a homogenizer at 900 bar; to form an acidified, disrupted microbial biomass;

b. heating the acidified, disrupted microbial biomass at 70° C. to form a heated, acidified, disrupted microbial biomass;

c. contacting the heated, acidified, disrupted microbial biomass with 1% DOWFAX2A1™ to release the one or more water immiscible compounds from the microbial biomass; and

d. recovering the one or more water immiscible compounds.

20. The method of claim 1 , comprising the steps of:

a. treating the microbial biomass by:

i. disrupting the microbial biomass in a homogenizer at 900 bar; and then

ii. acidifying the microbial biomass to a pH of 2.5; to form an acidified, disrupted microbial biomass;

b. heating the acidified, disrupted microbial biomass at 70° C. to form a heated, acidified, disrupted microbial biomass;

c. contacting the heated, acidified, disrupted microbial biomass with 1% DOWFAX2A1™ to release the one or more water immiscible compounds from the microbial biomass; and

d. recovering the one or more water immiscible compounds.

21. The method of claim 1 , wherein the microbial biomass has been de-emulsified with a de-emulsifying surfactant prior to step a.

22. The method of claim 21 , wherein the de-emulsifying surfactant is TERGITOL L-62™.

23. The method of claim 22 , wherein the amount of surfactant consumed is increased compared to the amount used in a comparable method lacking at least step c.

24. The method of claim 1 , wherein the microbial biomass is a recombinant yeast.

25. The method of claim 1 , wherein the microbial biomass is a recombinant Saccharomyces cerevisiae.

26. The method of claim 1 , wherein the microbial biomass comprises microbial cells that recombinantly express enzymes producing one or more water immiscible compounds.

27. The method of claim 1 , wherein the water immiscible compound is a C 5 to C 40 terpene, C 5 to C 20 terpene, or C 15 terpene.

28. The method of claim 1 , wherein the water immiscible compound is 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.

29. The method of claim 1 , wherein the water-immiscible compounds is α-farnesene or β-farnesene.