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Over the past decade, large efforts have been put into the metabolic engineering of cyanobacteria to produce various compounds such as alcohols, isoprenoids, biopolymers, and recombinant proteins. Forskolin is a structurally complex labdane-type diterpenoid with eight chiral carbon atoms and is naturally produced in the root cork of the plant Plectranthus barbatus. Forskolin is a potent cAMP activator indicated as a pharmaceutical for a variety of diseases. In the plant, forskolin biosynthesis from geranylgeranyl diphosphate involves six enzymes: two terpene synthases, three cytochrome P450s, and a single acetyltransferase. In this work, we express all six biosynthetic genes from Plectranthus barbatus in Synechocystis sp. PCC. 6803 and demonstrate heterologous production of this complex diterpenoid in a phototroph cyanobacterium. Forskolin titers reached 25.0 \u00b1 4.4 \u00b5g\/L and the forskolin was entirely secreted into the media. The forskolin-producing Synechocystis strain and empty vector control were cultivated in a photobioreactor for 8 days. Both strains showed similar chlorophyll a contents, and the forskolin-producing strain reached a slightly higher OD730 than the control. Forskolin began accumulating in the supernatant after 4 days and increased over time. These results indicate that forskolin production did not negatively impact cell growth.<\/jats:p>","DOI":"10.3390\/bioengineering12070683","type":"journal-article","created":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T08:50:57Z","timestamp":1750755057000},"page":"683","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Heterologous Production of the Structurally Complex Diterpenoid Forskolin in Synechocystis sp. PCC. 6803"],"prefix":"10.3390","volume":"12","author":[{"given":"Nadia","family":"Dodge","sequence":"first","affiliation":[{"name":"Department of Food Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lawrence Chuk","family":"Sutardja","sequence":"additional","affiliation":[{"name":"Section for Molecular Plant Biology, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3364-5804","authenticated-orcid":false,"given":"Silas","family":"Mellor","sequence":"additional","affiliation":[{"name":"Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thiyagarajan","family":"Gnanasekaran","sequence":"additional","affiliation":[{"name":"Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"L\u00e6rke Marie M\u00fcnter","family":"Lassen","sequence":"additional","affiliation":[{"name":"Section for Molecular Plant Biology, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Agnieszka Zygadlo","family":"Nielsen","sequence":"additional","affiliation":[{"name":"Section for Molecular Plant Biology, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3252-3119","authenticated-orcid":false,"given":"Birger Lindberg","family":"M\u00f8ller","sequence":"additional","affiliation":[{"name":"Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"},{"name":"VILLUM Center for Plant Plasticity, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6524-7723","authenticated-orcid":false,"given":"Poul Erik","family":"Jensen","sequence":"additional","affiliation":[{"name":"Department of Food Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg, Denmark"},{"name":"Section for Molecular Plant Biology, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Klaus, O., Hilgers, F., Nakielski AHasenklever, D., Jaeger, K.-E., Axmann, I.M., and Drepper, T. 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