Biological methods for preparing a fatty dicarboxylic acid
The technology relates in part to biological methods for producing a fatty dicarboxylic acid and engineered microorganisms capable of such production.
1. A method for producing a diacid, comprising:
contacting a genetically modified yeast with a feedstock capable of being converted by the yeast to a diacid; and
culturing the yeast under conditions in which the diacid is produced from the feedstock,
wherein the genetically modified yeast comprises: one or more genetic modifications that substantially block beta oxidation activity; and one or more genetic modifications that increase ADH2a alcohol dehydrogenase activity, wherein ADH2a alcohol dehydrogenase comprises the amino acid sequence of SEQ ID NO: 115.
2. The method of claim 1 , wherein the genetically modified yeast comprises one or more genetic modifications that increase a monooxygenase activity.
3. The method of claim 2 , wherein the monooxygenase comprises CYP52A12, CYP52A13, CYP52A14, CYP52A15, CYP52A16, CYP52A17, CYP52A18, CYP52A19 or CYP52A20.
4. The method of claim 1 , wherein the genetically modified yeast comprises one or more genetic modifications that increase a monooxygenase reductase activity.
5. The method of claim 4 , wherein the monooxygenase reductase is cytochrome p450 reductase B (CPRB).
6. The method of claim 1 , wherein the genetically modified yeast comprises one or more genetic modifications that disrupt activity of a POX4 polypeptide or a POX5 polypeptide.
7. The method of claim 1 , wherein the genetically modified yeast is a Candida spp. yeast.
8. The method of claim 7 , wherein the Candida spp. yeast is chosen from C. tropicalis and C. viswanathii.
9. The method of claim 7 , wherein the Candida spp. yeast is a genetically modified ATCC20336 yeast.
10. The method of claim 1 , wherein the feedstock comprises a substantially pure oil.
11. The method of claim 1 , wherein the feedstock comprises a plurality of fatty acids.
12. The method of claim 1 , wherein the feedstock comprises one or more components from a vegetable oil.
13. The method of claim 12 , wherein the vegetable oil is from a plant chosen from palm, palm kernel, coconut, soy, safflower, canola or combination thereof.
14. The method of claim 1 , wherein the diacid contains one or more unsaturations.
15. The method of claim 1 , wherein the diacid is a C8, C10, C12, C14, C16, C18 or C20 diacid.
16. The method of claim 15 , wherein the diacid is a C12 diacid.
17. The method of claim 1 , wherein the diacid is dodecanedioic acid or 12-hydroxydodecanoic acid.