# Biotechnological Fungal Platforms for the Production of Biosynthetic Cannabinoids
## 5 Conclusions  
The biosynthetic production of cannabinoids in fungal hosts presents great advantages in terms of the cost and productivity of therapeutic cannabinoids. This strategy could yield greater amounts of pure products with increased stability and robustness and at a lower cost in terms of the activities and resources required compared to those necessary for the  
Figure 4: Production of \(\Delta^{9}\)-THCA in _P. chrysogenum_ lysates after the addition of OA. (**a**) Schematic representation of the strategy followed to produce \(\Delta^{9}\)-THCA in _P. chrysogenum_ F-THCAS. Plasmid pSK-F-erg20-nphB included: _PpcbAB_-PpcbC (_P. chrysogenum_ pcbAB_-pcbC intergenic region), _TryrG_ (transcriptional terminator of the _pyrG_ gene), _erg20\({}^{\textit{699/N1227W}}\)_ (mutant variant of the GPP synthase gene), _PgpdA_ (promoter of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene from _A. nidulans_), cycl1 (transcriptional terminator of the _S. cerevisiae cycl1_ gene), _nphB\({}^{\textit{7288/A/G2865}}\)_ (mutant variant of the soluble aromatic phenyltransferase StNphB), and phleomycin resistance gene). (**b**) Chromatogram at 270 nm of one of the transformantsants of the F-THCAS-erg20-nphB strain showing the formation of \(\Delta^{9}\)-THCA (blue line). Chromatogram peaks corresponding to standards of cannabinoids are included (black line).  
traditional process of extraction from the plant or chemical synthesis. Yeasts have emerged as suitable platforms for the industrial production of cannabinoids, and the optimization of the production process can lead to increased titers. Based upon the experience of previous industrial processes for the production of secondary metabolites, filamentous fungi could also represent an alternative host for the cannabinoid industry. Since _P. chrysogenum_ has demonstrated its robustness under extreme industrial conditions in the production of penicillin, it could constitute a good candidate for the biosynthesis of other secondary metabolites. Herein, we provided evidence that this filamentous fungus could be genetically modified to produce small amounts \(\Delta^{9}\)-THCA after feeding cultures with CBGA or after the addition of OA to cell lysates, thereby confirming the feasibility of using this type of fungal host for this purpose. Although there is still much to be achieved in this field, including the optimization of expression cassettes, biosynthetic enzymes, and culture conditions and the redirection of metabolic fluxes, we believe that _P. chrysogenum_ could become a suitable biotechnological fungal platform for the production of cannabinoids in the near future.  
Conceptualization, C.G.-E.; methodology, C.G.-E., K.K., L.C. and I.-C.S.-O.; formal analysis, C.G.-E., K.K. and I.-C.S.-O.; investigation, C.G.-E., K.K., L.C. and I.-C.S.-O.; resources, C.G.-E. and C.B.; data curation, K.K. and I.-C.S.-O.; writing--original draft preparation, C.G.-E., K.K., L.C. and C.B., writing--review and editing, C.G.-E. and C.B.; visualization, C.G.-E. and L.C.; supervision, C.G.-E. and C.B.; project administration, C.G.-E. and C.B.; funding acquisition, C.G.-E. and C.B. All authors have read and agreed to the published version of the manuscript.  
This research was funded by the Spanish Ministry of Science, Innovation and Universities (Agencia Estatal de Investigacion) through the project CANNABISYN, grant number RTC-2017-6173-1 (Programa Estatal de Investigacion, Desarrollo e Innovacion orientada a los retos de la Sociedad).  
Not applicable.  
Data Availability Statement: Data are available on request.