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

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.

journal_name

Microbiologyopen

journal_title

MicrobiologyOpen

authors

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

doi

10.1002/mbo3.925

subject

Has Abstract

pub_date

2019-12-01 00:00:00

pages

e925

issue

12

issn

2045-8827

journal_volume

8

pub_type

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