Method for producing a polyhydroxyalkanoate using a microorganism belonging to the genus Cupriavidus
AssigneeKaneka Corporation
InventorsShunsuke Sato, Tetsuya Fujiki, Keiji Matsumoto
Life of this patent
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Abstract
The present invention provides a microorganism that synthesizes a high-molecular-weight PHA, and a method for producing a high-molecular-weight PHA, which have a productivity of at least 100 g/L. The provision is achieved by controlling the specific activity of a PHA synthase in cells of a microorganism that belongs to the genus Cupriavidus and is capable of producing a PHA, to 0.1 U/mg-protein at most. The microorganism and the method enable industrially efficient production of a PHA with a weight average molecular weight of at least 4,000,000.
Claims16
A microorganism belonging to the genus Cupriavidus which is capable of producing a polyhydroxyalkanoate with a weight average molecular weight of at least 4,000,000 at a level of at least 100 g/L of productivity, the microorganism comprising
- a gene encoding a polyhydroxyalkanoate synthase having at least one nucleotide sequence selected from the group consisting of the following nucleotide sequences (1) to (4):
- (1) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:9;
- (2) a nucleotide sequence encoding a polypeptide that has a sequence homology of at least 95% with the amino acid sequence of SEQ ID NO:9 and has a polyhydroxyalkanoate synthesis activity;
- (3) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:10;
- and
- (4) a nucleotide sequence encoding a polypeptide that has a sequence homology of at least 95% with the amino acid sequence of SEQ ID NO:10 and has a polyhydroxyalkanoate synthesis activity,
- wherein the gene encoding a polyhydroxyalkanoate synthase is controlled by at least one nucleotide sequence selected from the group consisting of the following nucleotide sequences (a) or (b) in such a way that a specific activity of the polyhydroxyalkanoate synthase in cells is 0.1 U/mg-protein at most:
- (a) the nucleotide sequence of SEQ ID NO:11;
- (b) a nucleotide sequence having a sequence homology of at least 95% with the nucleotide sequence of SEQ ID NO:11 and regulating gene transcription.
A method for producing a polyhydroxyalkanoate with a weight average molecular weight of at least 4,000,000, the method comprising
- culturing in a carbon source a microorganism belonging to the genus Cupriavidus which is capable of producing the polyhydroxyalkanoate with a weight average molecular weight of at least 4,000,000 at a level of at least 100 g/L of productivity, the microorganism comprising
- a gene encoding a polyhydroxyalkanoate synthase having at least one nucleotide sequence selected from the group consisting of the following nucleotide sequences (1) to (4):
- (1) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:9;
- (2) a nucleotide sequence encoding a polypeptide that has a sequence homology of at least 95% with the amino acid sequence of SEQ ID NO:9 and that has a polyhydroxyalkanoate synthesis activity;
- (3) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:10;
- and
- (4) a nucleotide sequence encoding a polypeptide that has a sequence homology of at least 95% with the amino acid sequence of SEQ ID NO:10 and that has a polyhydroxyalkanoate synthesis activity,
- wherein the gene encoding the polyhydroxyalkanoate synthase is controlled by at least one nucleotide sequence selected from the group consisting of the following nucleotide sequences (a) or (b) in such a way that a specific activity of the polyhydroxyalkanoate synthase in cells is 0.1 U/mg-protein at most:
- (a) the nucleotide sequence of SEQ ID NO:11;
- (b) a nucleotide sequence having a sequence homology of at least 95% with the nucleotide sequence of SEQ ID NO:11 and regulating gene transcription.
Dependencies are inferred from the claim text.