Transformed microorganism and method of producing polyhydroxyalkanoate
Provided is a transformed microorganism that has a polyhydroxyalkanoate synthase gene and in which expression of a minD gene is enhanced. Also provided is a transformed microorganism that has a polyhydroxyalkanoate synthase gene and in which expression of a minC gene and a minD gene is enhanced. In this transformed microorganism, expression of a minE gene may be enhanced or reduced. Also provided is a method of producing a PHA, the method including the step of culturing any of the transformed microorganisms in the presence of a carbon source.
1. A transformed microorganism, belonging to the genus Cupriavidus , and comprising a polyhydroxyalkanoate synthase gene, wherein expression of a minC gene and a minD gene is enhanced as compared to a wild strain of the microorganism,
wherein
the minC gene is a gene having a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, or a polypeptide comprising an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO: 1,
the minD gene is a gene having a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or a polypeptide comprising an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO: 2, and
the polyhydroxyalkanoate synthase gene is a gene having a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NOS: 4, 5, 6, 7, or 8, or a polypeptide having polyhydroxyalkanoate synthase activity and comprising an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NOS: 4, 5, 6, 7, or 8, and
a size of microbial cells of the transformed microorganism that accumulates polyhydroxyalkanoate is larger compared to a size of microbial cells of the wild strain that accumulates the polyhydroxyalkanoate, and the transformed microorganism accumulates larger-size particles of the polyhydroxyalkanoate compared to the wild strain.
2. The transformed microorganism according to claim 1 , wherein expression of a minE gene is enhanced as compared to a wild strain of the microorganism,
wherein the minE gene is a gene having a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 3, or a polypeptide comprising an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO: 3.
3. The transformed microorganism according to claim 2 , which is transformed Cupriavidus necator.
4. A method of producing a polyhydroxyalkanoate, comprising culturing the transformed microorganism of claim 2 in the presence of a carbon source.
5. The method according to claim 4 , wherein the carbon source comprises an oil or a fatty acid.
6. The method according to claim 4 , wherein the carbon source comprises a sugar.
7. The method according to claim 4 , wherein the carbon source contains comprises carbon dioxide.
8. The method according to claim 4 , wherein the polyhydroxyalkanoate is a copolymer of at least two hydroxy alkanoates.
9. The method according to claim 8 , wherein the polyhydroxyalkanoate is a copolymer comprising 3-hydroxyhexanoate as a monomer unit.
10. The method according to claim 9 , wherein the polyhydroxyalkanoate is a copolymer of 3-hydroxybutyrate and 3-hydroxyhexanoate.
11. The transformed microorganism according to claim 1 , which is transformed Cupriavidus necator.
12. A method of producing a polyhydroxyalkanoate, comprising culturing the transformed microorganism of claim 1 in the presence of a carbon source.
13. The method according to claim 12 , wherein the carbon source comprises an oil or a fatty acid.
14. The method according to claim 12 , wherein the carbon source comprises a sugar.
15. The method according to claim 12 , wherein the carbon source comprises carbon dioxide.
16. The method according to claim 12 , wherein the polyhydroxyalkanoate is a copolymer of at last two hydroxyalkanoates.
17. The method according to claim 16 , wherein the polyhydroxyalkanoate is a copolymer comprising 3-hydroxyhexanoate as a monomer unit.
18. The method according to claim 17 , wherein the polyhydroxyalkanoate is a copolymer of 3-hydroxybutyrate and 3-hydroxyhexanoate.
19. The transformed microorganism according to claim 1 , wherein the size of microbial cells of the transformed microorganism that accumulates the polyhydroxyalkanoate is at least 2.24 μm, and the size of polyhydroxyalkanoate particles that are accumulated by the transformed microorganism is at least 1.94 μm.
20. The transformed microorganism according to claim 2 , wherein the size of microbial cells of the transformed microorganism that accumulates the polyhydroxyalkanoate is at least 2.34 μm, and the size of polyhydroxyalkanoate particles that are accumulated by the transformed microorganism is at least 1.87 μm.