Methods for the biosynthesis of polyesters
View Patent ↗Coexpression of a polyhydroxyalkanoic acid synthase and a either a fatty acid:acyl-CoA transferase or an acyl-CoA synthetase in cells enables the biosynthesis of polyester materials. Plasmids, bacteria, materials, and methods for the preparation of polyesters are disclosed.
1. A recombinant nucleic acid segment encoding: a) a polyhydroxyalkanoic acid synthase protein; and b) a fatty acid:acyl-CoA transferase protein.
2. The nucleic acid segment of claim 1 , wherein the polyhydroxyalkanoic acid synthase protein is expressed from a nucleic acid molecule isolated from Alcaligenes eutrophus.
3. The nucleic acid segment of claim 2 , wherein the Alcaligenes eutrophus polyhydroxyalkanoic acid synthase protein is encoded by the Alcaligenes eutrophus phaC polyhydroxyalkanoic acid synthase structural gene.
4. The nucleic acid segment of claim 1 , wherein the fatty acid:acyl-coenzyme A transferase protein is a 4-hydroxybutyrate:acyl-CoA transferase protein.
5. The nucleic acid segment of claim 4 , wherein the 4-hydroxybutyrate:acyl-coenzyme A transferase protein is expressed from a nucleic acid molecule isolated from Clostridium kluyveri.
6. The nucleic acid segment of claim 5 , wherein the Clostridium kluyveri 4-hydroxybutyrate:acyl-coenzyme A transferase protein is encoded by the Clostridium kluyveri orfZ 4-hydroxybutyrate:acyl-coenzyme A transferase structural gene.
7. The nucleic acid segment of claim 1 , further comprising a promoter functional in bacterial cells.
8. The nucleic acid segment of claim 1 , wherein: a) The sequence encoding a polyhydroxyalkanoic acid synthase protein is operably linked to its native promoter; and b) the sequence encoding a fatty acid:acyl-coenzyme A transferase protein is operably linked to its native promoter.
9. The nucleic acid segment of claim 1 , further comprising a promoter heterologous to: a) the sequence encoding a polyhydroxyalkanoic acid synthase protein; and b) the sequence encoding a fatty acid:acyl-coenzyme A transferase protein.
10. A recombinant vector comprising the nucleic acid segment of claim 1 .
11. A recombinant vector comprising the nucleic acid segment of claim 9 .
12. The recombinant vector of claim 11 , further defined as vector pKSSE5.3.
13. The recombinant vector of claim 11 , further defined as vector pSKSE5.3.
14. A recombinant vector comprising: a) nucleic acid segment encoding a polyhydroxyalkanoic acid synthase protein; b) a nucleic acid segment encoding a fatty acid:acyl-coenzyme A transferase protein; and c) a promoter heterologous to the nucleic acid segment encoding a polyhydroxyalkanoic acid synthase protein, and heterologous to the nucleic acid segment encoding a fatty acid:acyl-coenzyme A transferase protein.
15. A recombinant cell for producing polyhydroxyalkanoate comprising a nucleic acid segment, the nucleic acid segment encoding:
a) a polyhydroxyalkanoic acid synthase protein; and
b) a fatty acid:acyl-coenzyme A transferase protein.
16. The cell of claim 15 , further defined as a plant cell, mammalian cell, insect cell, fungal cell, or bacterial cell.
17. The cell of claim 16 , wherein the cell is a plant cell.
18. The cell of claim 16 , wherein the cell is a bacterial cell.
19. The bacterial cell of claim 18 , wherein the bacterial cell is Escherichia coli.
20. The bacterial cell of claim 19 , wherein the bacterial cell is strain Escherichia coli XL 1-Blue.
21. The cell of claim 15 , wherein the polyhydroxyalkanoic acid synthase protein is expressed from a nucleic acid molecule isolated from Alcaligenes eutrophus.
22. The cell of claim 21 , wherein the Alcaligenes eutrophus polyhydroxyalkanoic acid synthase protein is encoded by the Alcaligenes eutrophus phaC polyhydroxyalkanoic acid synthase structural gene.
23. The cell of claim 15 , wherein the fatty acid:acyl-coenzyme A transferase protein is a 4-hydroxybutyrate:acyl-coenzyme A transferase protein.
24. The cell of claim 23 , wherein the 4-hydroxybutyrate:acyl-coenzyme A transferase protein expressed from a nucleic acid molecule isolated from Clostridium kluyveri.
25. The cell of claim 24 , wherein the Clostridium kluyveri 4-hydroxybutyrate:acyl-coenzyme A transferase protein is encoded by the Clostridium kluyveri orfZ 4-hydroxybutyrate:acyl-coenzyme A transferase structural gene.