Host cells and methods for producing isoprenyl alkanoates
The invention provides for a method of producing an isoprenyl alkanoate in a genetically modified host cell. In one embodiment, the method comprises culturing a genetically modified host cell which expresses an enzyme capable of catalyzing the esterification of an isoprenol and a straight-chain fatty acid, such as an alcohol acetyltransferase (AAT), wax ester synthase/diacylglycerol acyltransferase (WS/DGAT) or lipase, under a suitable condition so that the isoprenyl alkanoate is produced.
1. A method for producing an isoprenyl alkanoate in a genetically modified host cell, the method comprising:
(a) culturing a genetically modified host cell, wherein the genetically modified host cell comprises (i) an enzyme capable of catalyzing the esterification of a isoprenol and a straight-chain fatty acid, and (ii) one or more nucleic acid constructs encoding an isoprenyl pyrophosphate synthase and a pyrophosphase that is capable of hydrolyzing an isoprenyl diphosphate to an alcohol,
such that the culturing results in the genetically modified host cell producing an isoprenyl alkanoate; wherein the isoprenyl alkanoate is one selected from the group consisting of isoprenyl acetate, isoprenyl butyrate, isoprenyl hexanoate, isoprenyl octanoate, and isoprenyl decanoate.
2. The method of claim 1 , wherein the genetically modified host cell comprises a nucleic acid construct encoding the enzyme capable of catalyzing the esterification of an isoprenol and a fatty acid, and the culturing results in the expression of the enzyme.
3. The method of claim 1 , wherein the host cell comprises a single nucleic acid construct that encodes the the enzyme capable of catalyzing the esterification of the isoprenol and the straight-chain fatty acid, the isoprenyl pyrosphosphate synthase and the pyrophosphase.
4. The method of claim 2 , further comprising the step of: introducing the nucleic acid construct into the genetically modified host cell, wherein the introducing step is prior the culturing step.
5. The method of claim 1 , further comprising the step of:
(b) recovering the produced isoprenyl alkanoate, wherein the recovering step is concurrent or subsequent to the culturing step.
6. The method of claim 5 , further comprising the step of:
(c) hydrogenating the recovered isoprenyl alkanoate to produce a hydrogenated isoprenyl alkanoate, wherein the hydrogenating step is concurrent or subsequent to the recovering step;
such that part or all of the recovered isoprenyl alkanoate is hydrogenated.
7. The method of claim 1 , wherein the enzyme is an alcohol acetyltransferase (AAT), wax ester synthase (WS/DGAT) or lipase, or a homologous enzyme thereof.
8. The method of claim 1 , wherein the AAT is Saccharomyces cerevisiae AAT.
9. The method of claim 1 , wherein the WS/DGAT is Acinetobacter calcoaceticus WS/DGAT.
10. The method of claim 1 , wherein the isoprenyl acetate is isopentenyl acetate, geranyl acetate, or farnesyl acetate.
11. The method of claim 1 , wherein the isoprenyl butyrate is isopentenyl butyrate, geranyl butyrate, or farnesyl butyrate.
12. The method of claim 1 , wherein the isoprenyl hexanoate is isopentenyl hexanoate, geranyl hexanoate, or farnesyl hexanoate.
13. The method of claim 1 , wherein the isoprenyl octanoate is isopentenyl octanoate, geranyl octanoate, or farnesyl octanoate.
14. The method of claim 1 , wherein the isoprenyl decanoate is isopentenyl decanoate, geranyl decanoate, or farnesyl decanoate.
15. The method of claim 6 , wherein the hydrogenated isoprenyl alkanoate is one selected from the group consisting of isoprenyl acetate, isoprenyl butyrate, isoprenyl hexanoate, isoprenyl octanoate, and isoprenyl decanoate.
16. The method of claim 15 , wherein the isoprenyl acetate is isopentyl acetate, 3,7-dimethyloctyl acetate, or 3,7,11-trimethyldodecyl acetate.
17. The method of claim 15 , wherein the isoprenyl butyrate is isopentanyl butyrate, 3,7-dimethyloctanyl butyrate, or 3,7,11-trimethyldodecanyl butyrate.
18. The method of claim 15 , wherein the isoprenyl hexanoate esters is isopentanyl hexanoate, 3,7-dimethyloctanyl hexanoate, or 3,7,11-trimethyldodecanyl hexanoate.
19. The method of claim 15 , wherein the isoprenyl octanoate is isopentanyl octanoate, 3,7-dimethyloctanyl octanoate, or 3,7,11-trimethyldodecanyl octanoate.
20. The method of claim 15 , wherein the isoprenyl decanoate is isopentanol decanoate, 3,7-dimethyloctanyl decanoate, or 3,7,11-trimethyldodecanyl decanoate.
21. The method of claim 1 , wherein the host cell is a eubacteria.
22. The method of claim 21 , wherein the host cell is one selected from the Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsielia, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla, Synechococcus, Synechocystis , and Paracoccus taxonomical classes.
23. The method of claim 22 , wherein the host cell is Escherichia coli.
24. The method of claim 1 , wherein the host cell is an algal, fungal, insect or mammalian cell line.
25. The method of claim 24 , wherein the host cell is a yeast.
26. A genetically modified host cell comprising: (i) a nucleic acid construct encoding an enzyme capable of catalyzing the esterification of an isoprenol and a straight-chain fatty acid into an isoprenyl alkanoate, and (ii) one or more nucleic acid constructs encoding an isoprenyl pyrophosphate synthase and a pyrophosphase that is capable of hydrolyzing an isoprenyl diphosphate to an alcohol; wherein the isoprenyl alkanoate is one selected from the group consisting of isoprenyl acetate, isoprenyl butyrate, isoprenyl hexanoate, isoprenyl octanoate, and isoprenyl decanoate.
27. The host cell of claim 26 , wherein host cell prior to genetic modification does not produce the isoprenyl alkanoate.