3-HYDROXYPROPIONIC ACID AND OTHER ORGANIC COMPOUNDS

• US 20080076167A1
• Filed: 11/19/2007
• Published: 03/27/2008
• Est. Priority Date: 11/20/2000
• Status: Active Grant

• Current Assignee: Cargill, Inc. (Agricultural Services Division)
• Original Assignee: Cargill, Inc. (Agricultural Services Division)
• Inventors: GOKARN, Ravi, SELMER, Thorsten, SELIFONOVA, Olga, GORT, Steven, BUCKEL, Wolfgang, JESSEN, Holly
• Granted Patent: US 7,638,316 B2
• Time in Patent Office: Days
• US Class Current: 435/135
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/0004 Oxidoreductases (1.)
• CPC Class Codes: C12P 7/18 polyhydric
• CPC Class Codes: C12P 7/40 containing a carboxyl group including Peroxycarboxylic acids
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids
• CPC Class Codes: C12P 7/52 Propionic acid; Butyric acids
• CPC Class Codes: C12P 7/625 Polyesters of hydroxy carboxylic acids
• CPC Class Codes: Y02P 20/52 using catalysts, e.g. selective catalysts

Abstract

Methods and materials related to producing 3-HP as well as other organic compounds are disclosed. Specifically, isolated nucleic acids, polypeptides, host cells, and methods and materials for producing 3-HP and other organic compounds are disclosed.

Claims

1. A method of making 3-hydroxypropionic acid (3-HP), comprising:

culturing at least one cell comprising at least one exogenous nucleic acid molecule encoding at least one polypeptide that is capable of producing said 3-HP from phosphoenolpyruvate (PEP), pyruvate, or lactate under conditions such that said 3-HP is produced.

2. The method of claim 1, wherein 3-HP is made from PEP.

3. The method of claim 1, wherein 3-HP is made by a biosynthetic route that utilizes a β

-alanine intermediate, an acetyl-CoA intermediate, a malonyl-CoA intermediate or a lactate intermediate.

4. The method of claim 3, wherein 3-HP is made by a biosynthetic route that utilizes a β

-alanine intermediate.

5. The method of claim 1, wherein the at least one polypeptide that is capable of producing said 3-HP from PEP comprises an enzyme that can deaminate β

-alanine to yield malonate semialdehyde, and the cell can produce the 3-HP from the malonate semialdehyde.

6. The method according to claim 5, wherein the enzyme that can deaminate β

-alanine to yield malonate semialdehyde comprises an aminotransferase.

7. The method of claim 6, wherein the aminotransferase comprises a 4-aminobutyrate aminotransferase.

8. The method of claim 7, wherein the 4-aminobutyrate aminotransferase is a Clostridium acetobutycilicum 4 aminobutyrate aminotransferase.

9. The transformed cell of claim 8, wherein the Clostridium acetobutycilicum 4-aminobutyrate aminotransferase comprises a gabT sequence shown in GenBank Accession No. AE007654.

10. The method of claim 2, wherein the at least one polypeptide that is capable of producing said 3-HP from PEP comprises a 3-hydroxyisobutyrate dehydrogenase or a 3-HP dehydrogenase, and wherein the cell can make the 3-HP from malonate semialdehyde.

11. The method of claim 10, wherein the 3-hydroxyisobutyrate dehydrogenase is a P. aeruginosa 3-hydroxyisobutyrate dehydrogenase.

12. The method of claim 11, wherein the P. aeruginosa 3-hydroxyisobutyrate dehydrogenase comprises a mmsB sequence shown in GenBank Accession No. M84911.

13. The method of claim 2, wherein the at least one polypeptide that is capable of producing said 3-HP from PEP comprises a 4-aminobutyrate aminotransferase, and a 3-hydroxyisobutyrate dehydrogenase or a 3-HP dehydrogenase, wherein the cell can make 3-HP from malonate semialdehyde.

14. The method of claim 1, further comprising contacting the 3-HP with a peptide having lipase activity to make an ester of 3-HP.

15. The method of claim 1, further comprising contacting the 3-HP with a polypeptide comprising oxidoreductase activity, a polypeptide comprising reductase activity, a polypeptide comprising aldehyde dehydrogenase activity, and/or a polypeptide comprising alcohol dehydrogenase activity to make 1,3-propanediol.

16. The method of claim 4, wherein the β

-alanine is contacted with a peptide having 4-aminobutyrate aminotransferase activity to form malonate semialdehyde.

17. The method of claim 4, wherein malonate semialdehyde is contacted with a peptide having 3-hydroxyisobutyrate dehydrogenase or 3-HP dehydrogenase activity to make 3-HP.

18. The method of claim 1, wherein 3-HP is made from pyruvate, and the at least one polypeptide that is capable of producing said 3-HP from pyruvate comprises one or more of aspartate decarboxylase, aspartate aminotransferase, pyruvate carboxylase, and glutamate dehydrogenase.

19. The method of claim 1, wherein the cell is prokaryotic.

20. The method of claim 1, wherein the cell is a yeast cell, Lactobacillus cell, Lactococcus cell, Bacillus cell, or Escherichia cell.

21. A method for making 3-HP, comprising:

contacting β

-alanine with a peptide having 4-aminobutyrate aminotransferase activity to form malonate semialdehyde; and

contacting the malonate semialdehyde with a peptide having 3-hydroxyisobutyrate dehydrogenase or 3-HP dehydrogenase activity to make 3-HP.

22. The method of claim 21, further comprising:

contacting aspartate with a peptide having aspartate decarboxylase activity to make the β

-alanine.

23. The method of claim 22, further comprising:

contacting oxaloacetate with a peptide having aspartate aminotransferase activity to make the aspartate.

24. The method of claim 23, further comprising:

contacting PEP with a peptide having PEP carboxylase activity to make the oxaloacetate.

25. The method of claim 24, further comprising:

contacting pyruvate with a peptide having pyruvate carboxylase activity to make the oxaloacetate.