Designing microorganisms for heterologous biosynthesis of cannabinoids

Abstract

During the last decade, the use of medical Cannabis has expanded globally and legislation is getting more liberal in many countries, facilitating the research on cannabinoids. The unique interaction of cannabinoids with the human endocannabinoid system makes these compounds an interesting target to be studied as therapeutic agents for the treatment of several medical conditions. However, currently there are important limitations in the study, production and use of cannabinoids as pharmaceutical drugs. Besides the main constituent tetrahydrocannabinolic acid, the structurally related compound cannabidiol is of high interest as drug candidate. From the more than 100 known cannabinoids reported, most can only be extracted in very low amounts and their pharmacological profile has not been determined. Today, cannabinoids are isolated from the strictly regulated Cannabis plant, and the supply of compounds with sufficient quality is a major problem. Biotechnological production could be an attractive alternative mode of production. Herein, we explore the potential use of synthetic biology as an alternative strategy for synthesis of cannabinoids in heterologous hosts. We summarize the current knowledge surrounding cannabinoids biosynthesis and present a comprehensive description of the key steps of the genuine and artificial pathway, systems biotechnology needs and platform optimization.

Engineering Escherichia coli as a platform for the in vivo synthesis of prenylated aromatics

“…To form the unique PA structure, a critical enzyme, aromatic prenyltransferase (aPTase), catalyzes the condensation reaction between a prenyl chain and an aromatic ring to yield the PA (Figure ). As most aPTases from plants are integral membrane proteins (Carvalho, Hansen, Kayser, Carlsen, & Stehle, ), this key enzyme also represents a major challenge in the development of a microbial platform for the synthesis of PAs as its functional heterologous expression is required to actively catalyze the prenylation reaction. As such, many proof of concept studies reconstituting PAs biosynthetic pathways have been done in eukaryotic cells (yeast; Z. Li et al, ; Zirpel, Degenhardt, Martin, Kayser, & Stehle, ), and even these efforts have been limited to whole‐cell biotransformations with low product yields.…”

Distribution of cannabinoid synthase genes in non-Cannabis organisms

“…Biosynthesis of unique compounds such as cannabinoids, terpenes and flavonoids in non-Cannabis organisms is gaining much interest as an industry or the research. Attempts for synthesizing THC and CBD in yeast and microorganisms are actively researched (Carvalho et al 2017). Heterologous production of isoprenoid, a large family of secondary metabolites, has been successful in both yeast and E.coli (Paddon and Keasling 2014).…”

Current Perspectives on Biotechnological Cannabinoid Production in Plants

“…For the latter possibility, it was recently shown that the tetrahydrocannabinolic acid (THCA)forming enzyme [7] in combination with a soluble prenyltransferase can be simultaneously produced in Saccharomyces cerevisiae and Komagataella phaffii in their active form [8]. This marks an important step towards the total biosynthesis of cannabinoids in a microbial platform organism and was recently reviewed in detail elsewhere [9].…”