Biosynthesis of cannabinoids and cannabinoid precursors
AssigneeGinkgo Bioworks, Inc.
InventorsElena Brevnova, Dylan Alexander Carlin, Brian Carvalho, Gabriel Rodriguez
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Abstract
Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro.
Claims127
A host cell that comprises a heterologous polynucleotide encoding an acyl activating enzyme (AAE), wherein the host cell is capable of activating more of a short chain fatty acid in the presence of Coenzyme A (CoA) than a control host cell that does not comprise the heterologous polynucleotide encoding the AAE.
A host cell that comprises a heterologous polynucleotide encoding an acyl activating enzyme (AAE), wherein the AAE comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO:
- 7.
A host cell that comprises a heterologous polynucleotide encoding an acyl activating enzyme (AAE) and a polyketide synthase (PKS), wherein the AAE comprises an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO:
- 7 and wherein the PKS comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:
- 82.
A method of synthesizing butyryl-CoA comprising contacting butyrate and Coenzyme A (CoA) with an acyl activating enzyme (AAE), wherein the AAE comprises an amino acid sequence that is at least 90% identical to the sequence of any one of SEQ ID NOs:
- 2-28.
A method of producing divaric acid (DA) comprising contacting butyrate and Coenzyme A (CoA) with:
- (1) an acyl activating enzyme (AAE), a polyketide synthase (PKS), and a polyketide cyclase (PKC);
- or
- (2) an acyl activating enzyme (AAE) and a bifunctional PKS-PKC,
- wherein the AAE comprises an amino acid sequence that is at least 90% identical to the sequence of any one of SEQ ID NOs:
- 2-28.
A method of producing divarinol (DL) comprising contacting butyrate with an acyl activating enzyme (AAE) and a polyketide synthase (PKS), wherein the AAE comprises an amino acid sequence that is at least 90% identical to the sequence of any one of SEQ ID NOs:
- 2-28.
A method of producing a cannabinoid compound or a cannabinoid precursor comprising culturing a host cell in the presence of butyrate, wherein the host cell comprises a heterologous polynucleotide encoding an acyl activating enzyme (AAE), and wherein the host cell is capable of producing more divarinol in the presence of butyrate than a host cell that expresses a heterologous polynucleotide encoding an AAE that comprises the sequence of SEQ ID NO:
- 1.
A non-naturally occurring polynucleotide encoding an acyl activating enzyme (AAE), wherein the polynucleotide comprises a sequence that is at least 90% identical to the sequence of any one of SEQ ID NOs:
- 30-56.
A non-naturally occurring polynucleotide encoding an acyl activating enzyme (AAE), wherein the polynucleotide comprises a sequence that is at least 90% identical to the sequence of SEQ ID NO:
- 35.
A bioreactor for producing a cannabinoid compound or a cannabinoid precursor, wherein the bioreactor contains an acyl activating enzyme (AAE), and wherein the AAE comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs:
- 2-28.
Dependencies are inferred from the claim text.