Biosynthetic approach to combine the first steps of cardenolide formation in Saccharomyces cerevisiae

Rieck C, Geiger D, Munkert J, Messerschmidt K, Petersen J, Strasser J, Meitinger N, Kreis W (2019)

Publication Language: English

Publication Type: Journal article, Original article

Publication year: 2019

Journal

Article Number: e925

DOI: 10.1002/mbo3.925

Abstract

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.

Authors with CRIS profile

Involved external institutions

Universität Potsdam Germany (DE)

How to cite

APA:

Rieck, C., Geiger, D., Munkert, J., Messerschmidt, K., Petersen, J., Strasser, J.,... Kreis, W. (2019). Biosynthetic approach to combine the first steps of cardenolide formation in Saccharomyces cerevisiae. MicrobiologyOpen. https://dx.doi.org/10.1002/mbo3.925

MLA:

Rieck, Christoph, et al. "Biosynthetic approach to combine the first steps of cardenolide formation in Saccharomyces cerevisiae." MicrobiologyOpen (2019).

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