Compounds, compositions, and methods for recovering water-immiscible compounds from microbial biomass

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

a. acidifying the microbial biomass;

b. disrupting the microbial biomass;

c. heating the microbial biomass;

d. contacting the microbial biomass with a surfactant in an amount sufficient to release at least 30% of the one or more water immiscible compounds from the microbial biomass; and

e. 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 2-4.

6. The method of claim 1 , wherein the acidifying step is at a pH of 2.5±0.2.

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

8. 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.

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

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

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

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

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

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

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

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

16. The method of claim 1 , wherein the microbial biomass is contacted with 0.2 to 1.5% surfactant.

17. The method of claim 1 , wherein the microbial biomass is contacted with 1.0%±0.1% surfactant.

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

19. 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.

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

a. acidifying the microbial biomass to pH 2.5±0.2; and then

b. disrupting the microbial biomass in a homogenizer at 900 bar;

c. heating the acidified, disrupted microbial biomass at 70° C.;

d. contacting the microbial biomass with 1% DOWFAX™ 2A1 to release the one or more water immiscible compounds from the microbial biomass; and

e. 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 or step b.

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 d.

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

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

26. The method of claim 1 , wherein the microbial biomass comprises microbial cells that recombinantly express enzymes producing the 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 compound is α-farnesene or β-farnesene.

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

disrupting the microbial biomass in a homogenizer at 900 bar; and then acidifying the microbial biomass to a pH of 2.5±0.2;

heating the microbial biomass at 70° C.;

contacting the microbial biomass with 1% DOWFAX 2A1™ to release the one or more water immiscible compounds from the microbial biomass; and

recovering the one or more water immiscible compounds.