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 genetically modified yeast, comprising an active, modified endogenous acyl-coA Oxidase polypeptide, wherein the genetically modified yeast is capable of producing a diacid from a feedstock comprising one or more components from a vegetable oil, and wherein the acyl-coA Oxidase polypeptide comprises an amino acid modification at one or more amino acid positions chosen from residues 81, 82, 83, 84, 85, 86, 88, 93, 94, 95, 96, 98, 102, 284, 287, 290, 291, 292, 294, 295, 436, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, and 463 of SEQ ID NO: 32.
2. The genetically modified yeast of claim 1 , wherein the acyl-coA Oxidase polypeptide comprises at least one amino acid substitution.
3. The genetically modified yeast of claim 1 , comprising a genetic modification that reduces the activity of an enoyl coA isomerase polypeptide.
4. The genetically modified yeast of claim 3 , wherein the genetic modification disrupts a polynucleotide that encodes the enoyl coA isomerase polypeptide.
5. The genetically modifies yeast of claim 3 , wherein the enoyl coA isomerase polypeptide is a polypeptide native to a yeast, and wherein the yeast is a Candida spp. yeast.
6. The genetically modified yeast of claim 1 , wherein the diacid is a C4 to C24 diacid.
7. The genetically modified yeast of claim 1 , wherein the diacid is a C10, C12, C14, C16, C18, or C20 diacid.
8. The genetically modified yeast of claim 1 , wherein the diacid is a predominant diacid in a mixture of diacids.
9. The genetically modified yeast of claim 1 , wherein the feedstock comprises a plurality of fatty acids.
10. The genetically modified yeast of claim 1 , wherein the vegetable oil is from a plant chosen from palm, palm kernel, coconut, soy, safflower, canola, palm, palm kernel, and any combination thereof.
11. The genetically modified yeast of claim 7 , wherein the diacid is a C12 diacid.
12. The genetically modified yeast of claim 11 , wherein the diacid is dodecanedioic acid.
13. The genetically modified yeast of claim 7 , wherein the diacid is a C10 diacid.
14. The genetically modified yeast of claim 13 , wherein the diacid is sebacic acid.