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Metabolic Engineering of Synechocystis sp. PCC 6803 for Production of the Plant Diterpenoid Manoyl Oxide
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Molecular Biomimetics.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Molecular Biomimetics.
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2015 (English)In: ACS Synthetic Biology, ISSN 0065-0897, E-ISSN 2161-5063, Vol. 4, no 12, 1270-1278 p.Article in journal (Refereed) Published
Abstract [en]

Forskolin is a high value diterpenoid with a broad range of pharmaceutical applications, naturally found in root bark of the plant Coleus forskohlii. Because of its complex molecular structure, chemical synthesis of forskolin is not commercially attractive. Hence, the labor and resource intensive extraction and purification from C. forskohlii plants remains the current source of the compound. We have engineered the unicellular cyanobacterium Synechocystis sp. PCC 6803 to produce the forskolin precursor 13R-manoyl oxide (13R-MO), paving the way for light driven biotechnological production of this high value compound. In the course of this work, a new series of integrative vectors for use in Synechocystis was developed and used to create stable lines expressing chromosomally integrated CfTPS2 and CfTPS3, the enzymes responsible for the formation of 13R-MO in C. forskohlii. The engineered strains yielded production titers of up to 0.24 mg g(-1) DCW 13R-MO. To increase the yield, 13R-MO producing strains were further engineered by introduction of selected enzymes from C. forskohlii, improving the titer to 0.45 mg g(-1) DCW. This work forms a basis for further development of production of complex plant diterpenoids in cyanobacteria.

Place, publisher, year, edition, pages
2015. Vol. 4, no 12, 1270-1278 p.
Keyword [en]
Synechocystis; manoyl oxide; forskolin; diterpenoid; MEP-pathway; genetic tools
National Category
Biochemistry and Molecular Biology
URN: urn:nbn:se:uu:diva-259966DOI: 10.1021/acssynbio.5b00070ISI: 000366884700002PubMedID: 26133196OAI: oai:DiVA.org:uu-259966DiVA: diva2:845940
Swedish Energy AgencyKnut and Alice Wallenberg FoundationEU, European Research Council, ERC-2012-ADG_20120314EU, European Research Council, 323034
Available from: 2015-08-13 Created: 2015-08-13 Last updated: 2016-02-24Bibliographically approved

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