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.

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Author:Christoph Paul Kurt RieckORCiDGND, Daniel Geiger, Jennifer MunkertORCiDGND, Katrin MesserschmidtORCiDGND, Jan Petersen, Juliane StrasserGND, Nadine Meitinger, Wolfgang KreisORCiDGND
DOI:https://doi.org/10.1002/mbo3.925
ISSN:2045-8827
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=31436030
Parent Title (English):Microbiologyopen
Publisher:Wiley
Place of publication:Hoboken
Document Type:Article
Language:English
Date of first Publication:2019/08/22
Year of Completion:2019
Release Date:2020/11/20
Volume:8
Issue:12
Page Number:11
Organizational units:Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Peer Review:Referiert
Publication Way:Open Access
Open Access / Gold Open-Access
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