Microbial production of cannabinoids
AssigneeAmyris, Inc.
InventorsVictor F. HOLMES, John E. HUNG, Irina A. KORYAKINA, Andrew P. KLEIN, Yi XIONG, William E. DRAPER, Hailley S. WARBINGTON
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Abstract
The compositions and methods of the disclosure can be used to produce a cannabinoid in a host cell, such as a yeast cell. For example, the disclosure features host cells (e.g., yeast cells) modified to express one or more enzymes of a cannabinoid biosynthetic pathway, such as an acyl activating enzyme (AAE), a tetraketide synthase (TKS), a cannabigerolic acid synthase (CBGaS), and/or an olivetolic acid cyclase (OAC).
Claims16
1-270. (canceled)
A non-naturally occurring CBGaS enzyme:
- (a) having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:
- 55, SEQ ID NO:56, SEQ ID NO:57, or SEQ ID NO:58;
- (b) having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:
- 63, optionally wherein the CBGaS has one or more amino acid substitutions relative to the amino acid sequence of SEQ ID NO:
- 63 selected from I109T, F119L, S245L, S247Y, M270T, C280L, S295D, V314L, A324F, and S361I;
- or
- (c) having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO;
- 64, optionally wherein the CBGaS has one or more amino acid substitutions relative to the amino acid sequence of SEQ ID NO:
- 64 selected from M275S, M275T, T276C, T276F, K291H, V292Y, V292H, V292F, G310C, F314N, A331C, A331T, and A347I.
A host cell comprising one or more heterologous nucleic acids that each, independently, encode
- (a) an acyl activating enzyme (AAE) having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 1-5 and 7-24, and/or
- (b) a tetraketide synthase (TKS) having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 25 and 27-43, and/or
- (c) a cannabigerolic acid synthase (CBGaS) having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 55-58, 63, and 64, and/or
- (d) an olivetolic acid cyclase (OAC) having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 45-52.
A method of genetically modifying a host cell, the method comprising introducing into the host cell one or more heterologous nucleic acids that each, independently, encode
- (a) an AAE having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 1-5 and 7-24, and/or
- (b) a TKS having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 25 and 27-43, and/or
- (c) a CBGaS having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOS:
- 55-58, 63, and 64, and/or
- (d) an OAC having an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NO:
- 45-52.
Dependencies are inferred from the claim text.