title: Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii.
abstract: The heterologous expression of terpene synthases in microbial hosts has opened numerous possibilities for bioproduction of desirable metabolites. Photosynthetic microbial hosts present a sustainable alternative to traditional fermentative systems, using freely available (sun)light and carbon dioxide as inputs for bio-production. Here, we report the expression of a patchoulol synthase from Pogostemon cablin Benth in the model green microalga Chlamydomonas reinhardtii. The sesquiterpenoid patchoulol was produced from the alga and was used as a marker of sesquiterpenoid production capacity. A novel strategy for gene loading was employed and patchoulol was produced up to 922±242µgg-1CDW in six days. We additionally investigated the effect of carbon source on sesquiterpenoid productivity from C. reinhardtii in scale-up batch cultivations. It was determined that up to 1.03mgL-1sesquiterpenoid products could be produced in completely photoautotrophic conditions and that the alga exhibited altered sesquiterpenoid production metabolism related to carbon source.
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Lauersen
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Baier
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany.
authors.forename: Julian
authors.initials: J
authors.lastname: Wichmann
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany.
authors.forename: Robin
authors.initials: R
authors.lastname: Wördenweber
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Mussgnug
authors.affiliation: Biomolecular Photonics, Department of Physics, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hübner
authors.affiliation: Biomolecular Photonics, Department of Physics, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huser
authors.affiliation: Bielefeld University, Faculty of Biology, Center for Biotechnology (CeBiTec), Universitätsstrasse 27, 33615 Bielefeld, Germany. Electronic address: olaf.kruse@uni-bielefeld.de.
authors.forename: Olaf
authors.initials: O
authors.lastname: Kruse
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D016825:Chlamydomonas reinhardtii / Q000502:physiology* / Q000528:radiation effects; D024861:Genetic Enhancement / Q000379:methods*; D007535:Isomerases / Q000235:genetics* / Q000378:metabolism; D008027:Light; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D010788:Photosynthesis / Q000502:physiology* / Q000528:radiation effects; D012717:Sesquiterpenes / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D012717:Sesquiterpenes; D007535:Isomerases; C065995:patchoulol synthase; C052983:patchouli alcohol
keywords: Chlamydomonas reinhardtii; Microalgae; Patchoulol; Sesquiterpenoids; Terpenoids