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Enzymes Involved in Phytocannabinoid Biosynthesis or Cannabinoid Metabolism
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Phytocannabinoids are bioactive natural products found in some flowering plants, liverworts, and fungi that can be beneficial for the treatment of human ailments such as pain, anxiety, and cachexia. Targeted biosynthesis of cannabinoids with desirable properties requires identification of the underlying genes and their expression in a suitable hete...
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Citations
... As a result, research on cannabinoid-based medications for treating inflammation and pain has been rapidly growing, especially due to the opioid crisis (Bouchet and Ingram 2020), as well as the search for cannabinoids from other sources (Gülck and Møller 2020). Moreover, CB1 receptor has shown greater significance in this context compared to CB2 receptor (Ruhl et al. 2021), and PET diastereoisomer presents better pharmacological effect than 9-trans-THC due to ligation on CB1, reducing neuroinflammation through reduction of prostaglandin D2 and prostaglandin E2 (Chicca et al. 2018). ...
... In C. sativa, olivetolic acid is a precursor in the biosynthesis of cannabinoids, while in liverworts, this function is carried out by stilbene acid, a compound related to biotic and abiotic stresses (Teka et al. 2022). Lunularic acid (LA) is a stilbene derivative that acts in the biosynthetic pathway of bibenzyl compounds in liverworts, including Radula and Plagiochila (Hussain et al. 2019;Gülck and Møller 2020). This molecule was first isolated from the liverwort Lunularia cruciata (L.) Lindb. ...
Bryophytes, particularly liverworts, are known for producing bioactive compounds with therapeutic potential. In this review, we highlight the anti-inflammatory activities of liverworts, focusing on their ability to synthesize cannabinoids, compounds previously identified in only two genera. We explore the pharmacological parallels between liverwort cannabinoids and those from Cannabis sativa, emphasizing molecular similarities and interactions with the endocannabinoid system (ECS). Despite the promising nature of these compounds, there is a marked scarcity of studies exploring the pharmacological applications of liverwort-derived cannabinoids. This gap in research underscores the need for further investigation into their therapeutic potential. Additionally, we propose that cannabinoids or similar compounds may be more widespread in liverwort taxa than currently recognized, hypothesizing that other orders could also harbor these bioactive molecules. The potential discovery of new cannabinoids in liverworts could offer novel avenues for pharmacological exploitation. We conclude by calling for expanded chemical analyses to uncover more liverwort species with medicinally relevant compounds, which may reveal broader anti-inflammatory applications and therapeutic benefits.
... After the decarboxylation process, three neutral cannabinoids (tetrahydrocannabinol, THC; cannabidiol, CBD; and cannabichromene, CBC) are derived from acidic cannabinoid forms [1,23]. CBGAS is a prenyltransferase, whereas THCAS, CBDAS, and CBCAS are all closely related oxidocyclases [24]. Almost all cannabis plants contain an active CBGAS that converts precursor molecules into CBGA, which is then metabolized to form THCA or CBDA. ...
The diverse hormonal treatments applied to hemp (Cannabis sativa L.) carry significant implications for cultivation, and yield optimization across a range of applications, including fiber, seed, oil production, and the enhancement of medicinal compounds. However, there is no evidence concerning the long-term consequences of hormonal treatment. To determine the connection between the effects of hormonal treatment and cannabinoid accumulation, hemp plants were treated with γ-aminobutyric acid (GABA), abscisic acid (ABA), and salicylic acid (SA) to investigate their effects on gene expression and cannabinoid content levels in female inflorescences at 3 days and 4 weeks after treatment. The treatments influenced the transcript levels of five key cannabinoid biosynthesis genes, with 1.0 mM GABA significantly increasing OAC, THCAS, and CBCAS transcripts within 48 to 72 h. Additionally, 1.0 mM GABA led to a significant increase in tetrahydrocannabinol content by day three and significant increases in total cannabidiol and cannabinoid by week four. In addition, both ABA and SA induced transient, dose-dependent increases or decreases in gene expressions, but cannabinoid accumulation at 4 weeks showed no significant changes compared to the control. These results provide valuable insights for hormonal application in cultivation and the development of traits that enhance cannabinoid production in cannabis cultivation, which could significantly contribute to optimizing industrial applications.
... Despite its domestication and use of at least 4,000 years B.P. (Long et al., 2017;McPartland et al., 2018;Ren et al., 2021), Hemp (Cannabis sativa L.) has only recently gained notoriety for its unique characteristics. Among them, there is the production of exclusive secondary metabolites, the phytocannabinoids, such as cannabidiol (CBD) and delta-9-tetrahydrocannabinol (THC) (Gülck and Møller, 2020;Hanuš et al., 2016), which have several pharmacological applications (Bonini et al., 2018;Page et al., 2020). Glandular trichomes mainly synthesize these compounds in high density in the floral buds of the female plant (Kim and Mahlbeg, 1997;Stout et al., 2012;Gülck and Møller, 2020). ...
... Among them, there is the production of exclusive secondary metabolites, the phytocannabinoids, such as cannabidiol (CBD) and delta-9-tetrahydrocannabinol (THC) (Gülck and Møller, 2020;Hanuš et al., 2016), which have several pharmacological applications (Bonini et al., 2018;Page et al., 2020). Glandular trichomes mainly synthesize these compounds in high density in the floral buds of the female plant (Kim and Mahlbeg, 1997;Stout et al., 2012;Gülck and Møller, 2020). Other relevant products, with different properties, are terpenoids, phenylpropanoids, acyl sugars, and fiber (Andre et al., 2016). ...
Cannabis sativa has a variety of industrial interest products, such as phytocannabinoids, terpenoids, phenylpropanoids, acyl sugars, and fibers. Several described pathogens, including viral species, will impact the current green cannabis revolution. The recent sequencing of its genome and transcriptome, allowing the optimization and understanding of the production of the metabolite, are a relevant tool for viruses presence analyzing. Using the cannabis transcriptome and Data mining analysis, we describe the first amalgavirus infecting Cannabis, the Cannabis sativa amalgavirus 1 (CSA1). The plant amalgaviruses has nine species infecting relevant crops. Like the other genus members, this cannabis virus has approximately 3.5 kb with two partially overlapping putative open reading frames with the characteristic +1 programmed ribosomal frameshifting. Mainly detected in the male plant, the CSA1 mapped reads were present in the flower and leaf tissues. Nevertheless, the possible impacts of viral replication on host metabolism and the production of secondary compounds are unknown.
... Polyketide (PK) pathway is the synthesis pathway of cannabinoids that starts from activated hexanoic acid in the form of hexanoyl-CoA [19][20][21][22][23][24][25]. The first step of this pathway begins with a condensation between hexanoyl-CoA and malonyl-CoA residues to form a 12-carbon tetraketide intermediate, under the action of the enzyme tetraketide synthase (TKS). ...
Objectives/Background: The Cannabis genus contain a mixture of cannabinoids and other minor components which have been studied so far. In this narrative review, we highlight the main aspects of the polarized discussion between abuse and toxicity versus the benefits of the compounds found in the Cannabis sativa plant. Methods: We investigated databases such as PubMed, Google Scholar, Web of Science and World Anti-doping Agency (WADA) documents for scientific publications that can elucidate the heated discussion related to the negative aspects of addiction, organ damage and improved sports performance and the medical benefits, particularly in athletes, of some compounds that are promising as nutrients. Results: Scientific arguments bring forward the harmful effects of cannabinoids, ethical and legislative aspects of their usage as doping substances in sports. We present the synthesis and metabolism of the main cannabis compounds along with identification methods for routine anti-doping tests. Numerous other studies attest to the beneficial effects, which could bring a therapeutic advantage to athletes in case of injuries. These benefits recommend Cannabis sativa compounds as nutrients, as well as potential pharmacological agents. Conclusions and Future Perspectives: From the perspective of both athletes and illegal use investigators in sport, there are many interpretations, presented and discussed in this review. Despite many recent studies on cannabis species, there is very little research on the beneficial effects in active athletes, especially on large groups compared to placebo. These studies may complete the current vision of this topic and clarify the hypotheses launched as discussions in this review.
... To date, 125 cannabinoids (or phytocannabinoids) have been identified and classified into eleven cannabinoid sub-classes [1]. While phytocannabinoids were once thought to be exclusively isolated from Cannabis sativa, currently, it is known that they also occur in flowering plants, liverworts, and even fungi [2]. ...
... The major phytocannabinoids produced by Cannabis sativa are Tetrahydrocannabinol (THC) and Cannabidiol (CBD) [3], with THC as the main psychotropic molecule [1]. However, minor phytocannabinoids, such as Cannabigerol (CBG), Cannabigerolic acid (CBGA), Cannabidiolic acid (CBDA), Cannabichromene (CBC), and Cannabichromenic acid (CBCA), are also expressed in lower but significant abundance [2]. In opposition to THC, these are not psychotropic drugs, making their clinical use safer compared to THC. ...
... Phytocannabinoids are meroterpenoids, molecules with a resorcinyl core typically decorated with a para-positioned alkyl, aralkyl, or isoprenyl side chain [2]. In phytocannabinoids, usually one of the side chains is a propyl or pentyl [2,3] (Figure 1). ...
Cannabinoids are widely recognized for their potential therapeutic effects, making them significant and valuable candidates for medical research and applications across various fields. This review aims to analyze the pharmacokinetics of Cannabidiol (CBD), Cannabigerol (CBG), and Cannabichromene (CBC), along with their corresponding acidic forms, Cannabidiolic acid (CBDA), Cannabigerolic acid (CBGA), and Cannabichromenic acid (CBCA). Among these cannabinoids, CBD is the most extensively studied. Nevertheless, research involving all the mentioned cannabinoids has shown that their pharmacokinetic parameters are highly variable, depending significantly on factors such as dose, formulation, route of administration, and diet. Furthermore, challenges such as brain penetration and first-pass metabolism have been highlighted. In conclusion, this review demonstrates significant progress in understanding the pharmacokinetics of non-psychotropic cannabinoids. However, it also underscores the need for further research, particularly on CBG, CBC, and their respective acidic forms, with the most significant gap being in clinical investigations. Expanding these studies is essential to facilitate their optimized use in medical treatments.
... So far, more than 150 cannabinoids have been isolated and identified [2,3]. They can be classified into three main groups: acidic ones that are enzymatically biosynthesized by the plant; neutral ones that are directly decarboxylated from the acidic ones by heat, either in the plant or after harvest; and others, including both acidic and neutral ones, that are formed from other non-enzymatic processes, e.g., isomerization, decomposition, and/or degradation [2][3][4]. ...
A study was conducted to search for the best separation of eighteen cannabinoids, the maximum number of cannabinoids that have been quantified so far, for potency testing of hemp-based products using liquid chromatography diode array detector (LC-DAD). The investigation utilized four column types, all sharing the same dimension (150 mm × 2.1 mm) and core–shell particle size (2.7 µm), but different stationary phases: dimethyl-octadecyl (Poroshell 120 EC-C18), diisobutyl-octadecyl (Raptor ARC-18), reverse phase (RP)-carbamate (Cortecs Shield RP-18), and RP-amide (Ascentis Express RP-Amide). The resolution of adjacent cannabinoids was kept close to 1.5 or higher, while the separation time was kept as short as possible. The fastest separation was achieved within 15.0 min using two sequential Raptor ARC-18 columns, with a mobile phase consisting of 75.0% acetonitrile and 25.0% aqueous solution of 0.03% formic acid and 0.5 mM ammonium formate at pH 2.97, at a flow rate of 0.5 mL/min. A slightly improved resolution of the eighteen cannabinoids was obtained within 18.5 min using two sequential Poroshell 120 EC-C18 columns under similar conditions, except for a mobile phase containing 77.5% acetonitrile and a reduced flow rate of 0.45 mL/min due to backpressure higher than 600 bars. Furthermore, a rapid 7.0 min separation was achieved for potency testing of hemp-based products by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) using a Cortecs Shield RP-18 column, with a mobile phase consisting of 70.0% acetonitrile and 30.0% aqueous solution of 0.01% formic acid and 1 mM ammonium formate at pH 3.38 at a flow rate of 0.5 mL/min.
... The accumulation of PSMs in medical cannabis is influenced by various factors, including salinity (Formisano et al., 2024;Yep et al., 2020), water availability (Caplan et al., 2019;Morgan et al., 2024;Park et al., 2022), specific nutrient supply Caplan et al., 2017;Saloner and Bernstein, 2021, 2022a, 2022bShiponi and Bernstein, 2021;Song et al., 2023), and light quality (Ahsan et al., 2024;Brousseau et al., 2021;Danziger and Bernstein, 2021a;Holweg et al., 2024). Although PSM accumulation has been extensively studied, the responses of these compounds to variations in PPFD and air temperature remain less well explored (Gülck and Møller, 2020). Studies have shown contrasting effects of PPFD on PSM in various plant species (Downum et al., 1991;Nascimento and Fett-Neto, 2010;Ibrahim and Jaafar, 2012). ...
... Initially, PT1 was identified to catalyse CBGA biosynthesis (Gulck and Moller 2020), however recent reports have demonstrated that PT4 has higher enzymatic activity then PT1 and may be the enzyme responsible (Blatt-Janmaat and Qu 2021). However, PT4 was neither identified through our SWATH proteomic approach nor by previous shotgun proteomics of CsGTs (Conneely et al. 2021). ...
Key message
Cannabis trichome development progresses in distinct phases that underpin the dynamic biosynthesis of cannabinoids and terpenes.
Abstract
This study investigates the molecular mechanisms underlying cannabinoid and terpenoid biosynthesis in glandular trichomes of Cannabis sativa (CsGTs) throughout their development. Female Cannabis sativa c. Hindu Kush were cultivated under controlled conditions, and trichome development was analysed from week 3 to week 8 of the flowering period. We employed light microscopy, quantitative metabolomics and proteomics to analyse morphological changes in trichome secretory cell development, and temporal changes in metabolite accumulation and protein abundance. Our findings identified three distinct developmental phases: pre-secretory (T3), secretory (T6), and post-secretory (T8), the first time the three phases of trichome development have been identified and investigated in CsGTs. The pre-secretory phase was characterized by smaller secretory cells, limited metabolite accumulation and elevated levels of proteins involved in protein biosynthesis and cellular development. The secretory phase exhibited the highest biosynthetic activity, marked by larger secretory cells, increased plastidal activity, central carbon metabolism, and significant accumulation of cannabinoids and terpenoids. The post-secretory phase showed a decrease in secretory cell size, reduced metabolic activity, and a decrease in the abundance of primary and secondary metabolism enzymes, although THCA continued to accumulate. Key enzymes showed dynamic changes correlating with the stages of trichome development. This study provides a comprehensive understanding of the molecular mechanisms regulating cannabinoid and terpenoid biosynthesis in CsGTs, offering insights for enhancing the production of these valuable compounds through targeted breeding and biotechnological approaches.
... Cannabigerol (CBG) is a phytocannabinoid extracted from Cannabis sativa L. Phytocannabinoids are classified into over 113 types including CBG, cannabichromene (CBC), cannabidiol (CBD), Δ 9 -tetrahydrocannabinol (Δ 9 -THC), and cannabinol [9,10]. CBG is synthesized non-enzymatically by decarboxylating cannabigerolic acid (CBGA), the precursor molecule of other cannabinoids [11]. ...
Objectives
To determine growth inhibitory and anti-cancer effects of Cannabigerol (CBG) in human colorectal cancer cells.
Methods
Anti-proliferative effect of CBG was examined using MTT assay and two colorectal cancer cells (SW480 and LoVo cells). Cell death ratio was analyzed using Annexin V/PI staining experiment. Cell cycle distribution was analyzed using flow cytometry. We also performed western blot analysis on apoptotic marker proteins.
Results
CBG showed growth inhibitory effect in colorectal cancer cells using MTT assay. IC50 concentration of CBG was 34.89 μM in SW480 cells and 23.51 μM in LoVo cells. Annexin V/PI staining showed that CBG treatment increased apoptotic cells from 4.8% to 31.7% in SW480 cells and from 7.7% to 33.9% in LoVo cells. Flow cytometry confirmed that CBG increased sub G1 population via G1 arrest in both SW480 and LoVo cells. Western blot analysis showed that CBG increased expression levels of cell death-related proteins such as cleaved PARP-1, cleaved caspase 9, p53, and caspase 3.
Conclusion
CBG treatment shows antiproliferative activity and causes apoptosis of colorectal cancer cells, suggesting that CBG is applicable as a promising anticancer drug.
... Plant cannabinoids (phytocannabinoids) are terpenophenolic compounds exerting diverse biological effects in humans via the modulation of the endocannabinoid system (Ligresti et al., 2016). Originally thought to be exclusive to Cannabis sativa L. (Cannabaceae), they have now been discovered in other flowering plants, liverworts, and fungi, where their biosynthesis is thought to have arisen independently on multiple occasions (Gulck & Moeller, 2020). One example of this parallel evolution is in South African Helichrysum umbraculigerum (Asteraceae), which has yielded both C 5 alkyl (e.g. ...
... This report provoked much interest in Radula species as novel sources of medicinal compounds (Kumar et al., 2019;Gulck & Moeller, 2020;Arif et al., 2021). A review of Radula natural products world-wide (Asakawa et al., 2020) reported PET in Japanese R. campanigera and R. chinensis, and in Costa Rican R. laxiramea (Cullmann & Becker, 1999). ...
The potential of cannabinoids to address public health challenges has stimulated exploration into alternative sources and production technologies. Radula marginata, an endemic Aotearoa/New Zealand liverwort, produces the bibenzyl cannabinoid perrottetinene (PET), analogous to Cannabis psychoactive tetrahydrocannabinol (THC). Structural differences between PET and THC could alter therapeutic interactions and mitigate adverse side effects.
To understand the cannabinoid production potential of R. marginata, we analyzed 75 collections from three locations across several seasons, collaborating with kaitiaki Māori (indigenous guardians). Metabolic plasticity of the phytocannabinoids and plant growth was assessed under controlled growth conditions, and in in vitro culture.
Perrottetinene diol (trans‐PTD), analogous to cannabidiol (trans‐CBD), and its acid precursor (PTDA), were identified and fully characterized from nature for the first time. Bibenzyl‐4‐geranyl (BB4G), analogous to cannabigerol (CBG), and its corresponding acid (BB4GA), were also isolated. Radula marginata showed chemotypes dominated by PET, PTD, or BB4G, in striking analogy to the main Cannabis chemotypes. These site‐selective chemotypes persisted after growth under artificial lighting and in in vitro progeny, suggesting genetic control.
These results expand phytocannabinoid knowledge through the discovery of PTD analogous to CBD. They add a new dimension to liverwort cannabinoids and suggest convergent evolution of biosynthesis in two distant plant lineages.
