Biosynthesis of cannabigerol and cannabigerolic acid: the gateways to further cannabinoid production

Abstract

Cannabinoids are a therapeutically valuable class of secondary metabolites with a vast number of substituents. The native cannabinoid biosynthetic pathway of Cannabis sativa generates cannabigerolic acid, the common substrate to multiple cannabinoid synthases. The bioactive decarboxylated analogue of this compound, cannabigerol, represents an alternate gateway into the cannabinoid space either as a substrate to non-canonical cannabinoid synthase homologues or to synthetic chemical reactions. Herein, we describe the identification and repurposing of the aromatic prenyl transferase, AtaPT, which when coupled with native enzymes of C. sativa can form an E. coli production system for cannabigerolic acid in cell lysates, and cannabigerol in whole cells. Engineering of AtaPT, guided by structural analysis, was performed to enhance its kinetics towards cannabigerolic acid production for subsequent use in a proof-of-concept lysate system. For the first time, we show a synthetic biology platform for cannabigerol biosynthesis in E. coli cells by employing AtaPT under an optimised microbial system. Our results have therefore set the foundation for sustainable production of well researched and rarer cannabinoids in an E. coli chassis.

Advancement of Research Progress on Synthesis Mechanism of Cannabidiol (CBD)

“…It marks an important mileage for E. coli to produce cannabinoids. In 2023, Kearsey et al 125 realized the production of CBG in bacteria for the first time, demonstrating the potential of the microbial pathway to functional cannabinoids (Table 1). Although significant progress has been made in the biosynthesis of cannabinoids in microorganisms such as S. cerevisiae, achieving high CBD production is still a major challenge.…”

Engineering cannabinoid production in Saccharomyces cerevisiae

“…[32] In a recently published study, E. coli was engineered to produce CBG-C5 and CBGA-C5 using the aromatic prenyltransferase AtaPT from Aspergillus terreus. [33] However, heterologous production of cannabinoids in E. coli is likely limited to the production of CBGA-C5 and CBG-C5, as the expression of THCAS in a bacterial host system is challenging due to missing posttranslational modification. [34] Future potential of cannabinoid production in heterologous systems is demonstrated by several reports on the production of the precursor OA.…”