Biological methods for preparing adipic acid
The technology relates in part to biological methods for producing adipic acid and engineered microorganisms capable of such production.
1. A method for producing adipic acid, comprising:
(a) contacting a Candida yeast with a feedstock comprising a vegetable oil, wherein the Candida yeast comprises:
(i) a genetic alteration that partially blocks beta oxidation activity of acyl-CoA oxidase polypeptide,
(ii) a genetic alteration that reduces or eliminates long chain acyl-CoA synthetase activity, and
(iii) a genetic alteration that reduces or eliminates acyl-CoA synthetase activity,
wherein the genetic alteration of (i) reduces the amount of a polypeptide having an acyl-CoA oxidase activity, and the polypeptide is a POX4 polypeptide or a POX5 polypeptide;
(b) culturing the Candida yeast under conditions in which adipic acid is produced.
2. The method of claim 1 , wherein the adipic acid is produced at a level of at least 2 grams per liter of culture medium.
3. The method of claim 1 , wherein the adipic acid is produced at a level of at least 20% of maximum theoretical yield for the feedstock.
4. The method of claim 1 , wherein the vegetable oil is a palm oil or a soybean oil.
5. The method of claim 1 , wherein the vegetable oil is oleic acid.
6. The method of claim 1 , wherein the vegetable oil is a fatty acid soap stock.
7. The method of claim 1 , wherein the polypeptide is a POX4 polypeptide.
8. The method of claim 7 , wherein the Candida yeast comprises a genetic alteration that increases POX5 activity.
9. The method of claim 1 , wherein the Candida yeast is a Candida tropicalis yeast.
10. The method of claim 1 , wherein the genetic alteration of (ii) eliminates the long chain acyl-CoA synthetase activity.
11. The method of claim 10 , wherein the genetic alteration of (ii) comprises disrupting a polynucleotide that encodes an acyl-CoA synthetase (FAT1) polypeptide.
12. The method of claim 11 , wherein the FAT1 polypeptide comprises the amino acid sequence of SEQ ID NO: 51.
13. The method of claim 1 , wherein the genetic alteration of (iii) eliminates the acyl-CoA synthetase activity.
14. The method of claim 13 , wherein the genetic alteration of (iii) comprises disrupting a polynucleotide that encodes a ACS1 polypeptide.
15. The method of claim 14 , wherein the ACS1 polypeptide comprises the amino acid sequence of SEQ ID NO: 49.
16. The method of claim 1 , wherein the Candida yeast comprises a genetic alteration that increases a CYP52A15 activity.
17. The method of claim 1 , wherein the Candida yeast comprises a genetic alteration that increases a CYP52A16 activity.
18. The method of claim 1 , wherein the Candida yeast comprises a genetic alteration that increases a CYP52A19 activity.