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US 12,031,123 B2Granted
Recombinant bacterial cells and methods for producing poly(3-hydroxybutyrate-co-3-hydroxyvalerate)
AssigneeGenecis Bioindustries Inc.
InventorAdam William Westbrook
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Abstract
The disclosure provides recombinant bacterial host cells that metabolize and convert glycerol or volatile fatty acids (VFAs) to poly(3-hydroxybutyrate-co-3-hydroxyvalerate) or PHBV. The disclosure further provides methods of producing PHBV using the recombinant bacteria disclosed herein.
Claims32
A bacterial host cell, comprising the following nucleic acid molecules integrated into the bacterial host cell genome:
- a first operon comprising (a) a nucleic acid molecule encoding a PhaC protein, wherein the PhaC protein is a Cupriavidus sp. S-6 PhaC protein, (b) a nucleic acid molecule encoding a PhaA protein, wherein the PhaA protein is a Cupriavidus sp. S-6 PhaA protein, and (c) a nucleic acid molecule encoding a PhaB protein, wherein the PhaB protein is a Cupriavidus sp. S-6 PhaB protein, wherein the first operon comprises a first promoter;
- a second operon comprising:
- (i) a nucleic acid molecule encoding a BktB protein, wherein the BktB protein is a Cupriavidus gilardii QJ1 BktB protein, and (ii) a nucleic acid molecule encoding a PhaB protein, wherein the PhaB protein is a Cupriavidus sp. S-6 PhaB protein, wherein the second operon comprises a second promoter;
- a third operon, comprising:
- (a) a nucleic acid molecule encoding a FadE protein, and (b) a nucleic acid molecule encoding a FadB protein, wherein the third operon comprises a third promoter;
- and
- a fourth operon, comprising:
- (a) a nucleic acid molecule encoding a LvaE protein, wherein the LvaE protein is a Pseudomonas putida LvaE protein, and (b) a nucleic acid molecule encoding a propionate-CoA transferase, wherein the propionate-CoA transferase is a Clostridium propionicum propionate-CoA transferase (Pct(Cp)), wherein the fourth operon comprises a fourth promoter;
- wherein the bacterial host cell comprises an activated sleeping beauty mutase (Sbm) pathway.
6 more elements
A bacterial host cell, comprising:
- a first operon comprising (a) a nucleic acid molecule encoding a PhaC protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 250, (b) a nucleic acid molecule encoding a PhaA protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 248, (c) a nucleic acid molecule encoding a PhaB protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 249;
- a second operon comprising:
- (i) a nucleic acid molecule encoding a BktB protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 251, and (ii) a nucleic acid molecule encoding a PhaB protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 249;
- a third operon, comprising:
- (a) a nucleic acid molecule encoding a FadE protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 72, and (b) a nucleic acid molecule encoding a FadB protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 71;
- a fourth operon, comprising:
- (a) a nucleic acid molecule encoding a LvaE protein, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 253 and (b) a nucleic acid molecule encoding a propionate CoA-transferase, wherein the nucleic acid molecule comprises a sequence having at least 80% identity to SEQ ID NO:
- 89, and
- a sleeping beauty mutase (Sbm) operon comprises a Ptrc promoter,
- wherein each of the first, second and fourth operons comprises a promoter comprising the nucleic acid sequence of SEQ ID NO:
- 233 (Pgracmax2), and the third operon comprises a promoter comprising the nucleic acid sequence of SEQ ID NO:
- 254 (Ptrc).
17 more elements
Dependencies are inferred from the claim text.