Engineering of Saccharomyces cerevisiae for the production of (+)-ambrein

Sandra Moser, Erich Leitner, Thomas J Plocek, Koenraad Vanhessche, Harald Pichler

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Abstract

The triterpenoid (+)‐ambrein is the major component of ambergris, a coprolite of the sperm whale that can only be rarely found on shores. Upon oxidative degradation of (+)‐ambrein, several fragrance molecules are formed, amongst them (−)‐ambrox, one of the highest valued compounds in the perfume industry. In order to generate a Saccharomyces cerevisiae whole‐cell biocatalyst for the production of (+)‐ambrein, intracellular supply of the squalene was enhanced by overexpression of two central enzymes in the mevalonate and sterol biosynthesis pathway, namely the N‐ terminally truncated 3‐hydroxy‐3‐methylglutaryl‐CoA reductase 1 (tHMG) and the squalene synthase (ERG9). In addition, another key enzyme in sterol biosynthesis, squa- lene epoxidase (ERG1) was inhibited by an experimentally defined amount of the inhib- itor terbinafine in order to reduce flux of squalene towards ergosterol biosynthesis while retaining sufficient activity to maintain cell viability and growth. Heterologous expression of a promiscuous variant of Bacillus megaterium tetraprenyl‐β‐curcumene cyclase (BmeTC‐D373C), which has been shown to be able to catalyse the conversion of squalene to 3‐deoxyachillol and then further to (+)‐ambrein resulted in production of these triterpenoids in S. cerevisiae for the first time. Triterpenoid yields are comparable with the best microbial production chassis described in literature so far, the methylotrophic yeast Pichia pastoris. Consequently, we discuss similarities and differ- ences of these two yeast species when applied for whole‐cell (+)‐ambrein production.
Originalspracheenglisch
Seiten (von - bis)163 - 172
Seitenumfang10
FachzeitschriftYeast
Jahrgang37
Ausgabenummer1
DOIs
PublikationsstatusVeröffentlicht - 15 Feb 2020

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Schlagwörter

    ASJC Scopus subject areas

    • !!Biochemistry, Genetics and Molecular Biology(all)

    Fields of Expertise

    • Human- & Biotechnology

    Treatment code (Nähere Zuordnung)

    • Application
    • Basic - Fundamental (Grundlagenforschung)
    • Experimental

    Kooperationen

    • BioTechMed-Graz
    • NAWI Graz

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