Abstract: Provided herein are cells, enzymes, and methods for improved cannabinoid production.

Abstract: Described herein are the discovery and/or optimization of enzymes involved in the biosynthesis of olivetolic acid, divarinic acid, and analogs thereof.

Abstract: Disclosed herein are novel CBGA and CBGVA synthases and methods for improvement of their overall activities for the synthesis of CBGA and CBGVA from their respective precursors, olivetolic acid (OA) or divarinic acid (DVA) and GPP. Also disclosed are fusion proteins to enhance synthesis of CBGA and CBGVA. The methods described herein also increase the titer and the purity of CBGA and CBGVA made by a cell by 1) decreasing the formation of byproducts FCBGA and FCBGVA that are synthesized from the respective prenylation of OA and DVA with FPP, and/or 2) increasing the intracellular availability of OA and DVA.

Abstract: The invention provides an engineered carboxylic acid reductase (CAR) enzyme, a nucleic acid encoding the CAR enzyme, and a non-naturally occurring microbial organism comprising an exogenous nucleic acid encoding the CAR, an engineered transaminase (TA) enzyme, and/or a hexamethylenediamine (HMD) transaminase (TA2) enzyme. The invention provides a non-naturally occurring microbial organism that has a 1,6-hexanediol (HDO) pathway with a HDO pathway enzyme expressed in sufficient amounts to produce 6 aminocaproate semi aldehyde, HDO, or both. The invention further provides a non-naturally occurring microbial organism that has an HMD pathway with a HMD pathway enzyme expressed in sufficient amounts to produce 6-aminocaproate semialdehyde, HMD, or both. The invention additionally provides bioderived HMD, 6-aminocaproate semialdehyde, and/or HDO and methods for producing bioderived HMD, 6-aminocaproate semialdehyde, and/or HDO.

Abstract: Disclosed are transaminase (TA) enzymes and nucleic acids encoding them. In some cases, the transaminase enzymes are non-natural, engineered transaminases. Also disclosed are biosynthetic methods and engineered microorganisms that enhance or improve the biosynthesis of 6-aminocaproate, hexamethylenediamine, caproic acid, caprolactone, or caprolactam. The engineered microorganisms include exogenous TA and in some cases engineered TA.

Abstract: Described herein are olivetolic acid cyclases (OAC) including non-natural variants capable of forming a 2,4-dihydroxy-6-alkylbenzoic acid from a 3,5,7-trioxoacyl-CoA or a 3,5,7-trioxocarboxylate substrate. In some examples, the non-natural OAC is capable of forming a 2,4-dihydroxy-6-alkylbenzoic acid from a 3,5,7-trioxoacyl-CoA or a 3,5,7-trioxocarboxylate substrate at a greater rate. In some examples, the non-natural OAC has a higher affinity for a 3,5,7-trioxoacyl-CoA or a 3,5,7-trioxocarboxylate substrate, as compared to the wild type OAC. The non-natural OAC can be used with olivetol synthase (OLS) to form the 2,4-dihydroxy-6-alkylbenzoic acid from malonyl-CoA and acyl-CoA through to a 3,5,7-trioxoacyl-CoAintermediate. The non-natural OAC (and OLS) can be expressed in an engineered cell having a pathway to form cannabinoids, which include CBGA, its analogs and derivatives. CBGA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.

Abstract: Described herein are non-natural olivetol synthase (OLS) variants, nucleic acids, engineered cells, method s for preparing cannabinoids, and compositions thereof. The non-natural olivetol OLS variants form desired cannabinoid precursor and products at increased rates, have higher affinity for pathway substrates, and/or byproducts are formed in lower amounts in their presence, as compared to wild type OLS. The OLS variants can be used to form linear polyketides, and can be expressed in an engineered cell having a pathway to form cannabinoids, which include CBGA, its analogs and derivatives. CBGA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.