Microbial production of cannabinoids
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).
1 . A non-naturally occurring olivetolic acid cyclase (OAC) enzyme having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 44, wherein the OAC has one or more amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 44 selected from A2S, L9I, K12S, E14S, V28L, T47R, Q48R, K49R, S87H, F88Y, and L92Y.
2 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution A2S relative to the amino acid sequence of SEQ ID NO: 44.
3 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution L91 relative to the amino acid sequence of SEQ ID NO: 44.
4 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution K12S relative to the amino acid sequence of SEQ ID NO: 44.
5 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution E14S relative to the amino acid sequence of SEQ ID NO: 44.
6 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution F23L relative to the amino acid sequence of SEQ ID NO: 44.
7 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution V28L relative to the amino acid sequence of SEQ ID NO: 44.
8 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution T47R relative to the amino acid sequence of SEQ ID NO: 44.
9 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution Q48R relative to the amino acid sequence of SEQ ID NO: 44.
10 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution K49R relative to the amino acid sequence of SEQ ID NO: 44.
11 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution S87H relative to the amino acid sequence of SEQ ID NO: 44.
12 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution F88Y relative to the amino acid sequence of SEQ ID NO: 44.
13 . The non-naturally occurring OAC enzyme of claim 1 , wherein the OAC has the amino acid substitution L92Y relative to the amino acid sequence of SEQ ID NO: 44.
14 . A non-naturally occurring OAC enzyme 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.
15 . The non-naturally occurring OAC enzyme of claim 14 , wherein the OAC enzyme has an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOS: 45-52.
16 . The non-naturally occurring OAC enzyme of claim 14 , wherein the OAC enzyme has an amino acid sequence that comprises the amino acid sequence of any one of SEQ ID NOS: 45-52.
17 . The non-naturally occurring OAC enzyme of claim 14 , wherein the OAC enzyme has an amino acid sequence that comprises the amino acid sequence of SEQ ID NO: 45.
18 . The non-naturally occurring OAC enzyme of claim 1 , wherein the non-naturally occurring OAC enzyme is in a yeast cell.