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US 6,913,911 B2Granted
Transgenic microbial polyhydroxyalkanoate producers
AssigneeMetabolix, Inc.
InventorsGjalt W. Huisman, Oliver P. Peoples, Frank A. Skraly
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AssigneeMetabolix, Inc.
InventorsGjalt W. Huisman, Oliver P. Peoples, Frank A. Skraly
Estimating the term…
Transgenic microbial strains are provided which contain the genes required for PHA formation integrated on the chromosome. The strains are advantageous in PHA production processes, because (1) no plasmids need to be maintained, generally obviating the required use of antibiotics or other stabilizing pressures, and (2) no plasmid loss occurs, thereby stabilizing the number of gene copies per cell throughout the fermentation process, resulting in homogeneous PHA product formation throughout the production process. Genes are integrated using standard techniques, preferably transposon mutagenesis. In a preferred embodiment wherein mutiple genes are incorporated, these are incorporated as an operon. Sequences are used to stabilize mRNA, to induce expression as a function of culture conditions (such as phosphate concentration), temperature, and stress, and to aid in selection, through the incorporation of selection markers such as markers conferring antibiotic resistance.
A genetically engineered microorganism producing polyhydroxyalkanoate, the improvement comprising having at least one heterologous bacterial gene encoding enzyme involved in the synthesis of polyhydroxalkanoates selected from the group consisting of thiolase, reductase, PHB synthase, PHA synthase, acyl-CoA transferase, and enoyl-CoA hydratase integrated into the chromosome, which stably expresses the enzyme and thereby has enhanced production of polyhydroxyalkanoate.
A method for screening for a bacterial gene involved in synthesis of polyhydroxyalkanoates that enhances production comprising
A method for producing polyhydroxyalkanoates comprising culturing genetically engineered microorganisms capable of producing polyhydroxyalkanoates having at least one bacterial gene encoding an enzyme involved in synthesis of polyhydroxyalkanoates selected from the group consisting of thiolase, reductas, PHB synthase, PHA synthase, acyl-CoA transferase, enoyl-CoA hydratase, integrated into the chromosome, with appropriate substrate under conditions wherein the microorganisms produce polyhydroxyalkanoate.
Dependencies are inferred from the claim text.