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 disruption, deletion or knockout of (1) a polynucleotide that encodes a POX4 polypeptide or (2) a promoter operably linked to a polynucleotide that encodes a POX4 polypeptide, whereby POX4 activity is reduced or removed and POX5 activity is not reduced or removed,
(ii) a disruption, deletion or knockout of (1) a polynucleotide that encodes a long chain acyl-CoA synthetase polypeptide or (2) a promoter operably linked to a polynucleotide that encodes a long chain acyl-CoA synthetase polypeptide, whereby a long chain acyl-CoA synthetase activity is reduced or removed, and
(iii) a disruption, deletion or knockout of (1) a polynucleotide that encodes a long chain acyl-CoA synthetase polypeptide or (2) a promoter operably linked to a polynucleotide that encodes a long chain acyl-CoA synthetase polypeptide, whereby an acyl-CoA synthetase activity is reduced or removed; and
(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 Candida yeast comprises a genetic alteration that increases POX5 activity.
8. The method of claim 1 , wherein the Candida yeast is a genetically modified Candida strain ATCC20692.
9. The method of claim 1 , wherein the long chain acyl-CoA synthetase activity is removed.
10. The method of claim 9 , wherein the long chain acyl-CoA synthetase polypeptide is a FAT1 polypeptide.
11. The method of claim 10 , wherein the FAT1 polypeptide comprises the amino acid sequence of SEQ ID NO: 51.
12. The method of claim 1 , wherein the acyl-CoA synthetase activity is removed.
13. The method of claim 12 , wherein the acyl-CoA synthetase polypeptide is an ACS1 polypeptide.
14. The method of claim 13 , wherein the ACS1 polypeptide comprises the amino acid sequence of SEQ ID NO: 49.
15. The method of claim 1 , wherein the Candida yeast comprises a genetic alteration that increases the number of copies of a polynucleotide that encodes a CYP52A15, CYP52A16 or CYP52A19 polypeptide.
16. The method of claim 1 , wherein the Candida yeast comprises a genetic alteration that inserts a promoter in functional connection with a polynucleotide that encodes a CYP52A15, CYP52A16 or CYP52A19 polypeptide.
17. The method of claim 15 , wherein the Candida yeast comprises a genetic alteration that increases the number of copies of a polynucleotide that encodes a CYP52A19 polypeptide.
18. The method of claim 17 , wherein the CYP52A19 polypeptide is a Candida yeast polypeptide.
19. The method of claim 16 , wherein the Candida yeast comprises a genetic alteration that inserts a promoter in functional connection with a polynucleotide that encodes a CYP52A19 polypeptide.
20. The method of claim 19 , wherein the CYP52A19 polypeptide is a Candida yeast polypeptide.