Biosynthetic approach to combine the first steps of cardenolide formation in Saccharomyces cerevisiae
- A yeast expression plasmid was constructed containing a cardenolide biosynthetic module, referred to as CARD II, using the AssemblX toolkit, which enables the assembly of large DNA constructs. The genes cloned into the vector were (a) a Δ5‐3β‐hydroxysteroid dehydrogenase gene from Digitalis lanata, (b) a steroid Δ5‐isomerase gene from Comamonas testosteronii, (c) a mutated steroid‐5β‐reductase gene from Arabidopsis thaliana, and (d) a steroid 21‐hydroxylase gene from Mus musculus. A second plasmid bearing an ADR/ADX fusion gene from Bos taurus was also constructed. A Saccharomyces cerevisiae strain bearing these two plasmids was generated. This strain, termed “CARD II yeast”, was capable of producing 5β‐pregnane‐3β,21‐diol‐20‐one, a central intermediate in 5β‐cardenolide biosynthesis, starting from pregnenolone which was added to the culture medium. Using this approach, five consecutive steps in cardenolide biosynthesis were realized in baker's yeast.
Verfasserangaben: | Christoph Paul Kurt RieckORCiDGND, Daniel Geiger, Jennifer MunkertORCiDGND, Katrin MesserschmidtORCiDGND, Jan Petersen, Juliane StrasserGND, Nadine Meitinger, Wolfgang KreisORCiDGND |
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DOI: | https://doi.org/10.1002/mbo3.925 |
ISSN: | 2045-8827 |
Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/31436030 |
Titel des übergeordneten Werks (Englisch): | Microbiologyopen |
Verlag: | Wiley |
Verlagsort: | Hoboken |
Publikationstyp: | Wissenschaftlicher Artikel |
Sprache: | Englisch |
Datum der Erstveröffentlichung: | 22.08.2019 |
Erscheinungsjahr: | 2019 |
Datum der Freischaltung: | 20.11.2020 |
Band: | 8 |
Ausgabe: | 12 |
Seitenanzahl: | 11 |
Organisationseinheiten: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
DDC-Klassifikation: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
Peer Review: | Referiert |
Publikationsweg: | Open Access |
Open Access / Gold Open-Access | |
DOAJ gelistet |