BioChem - DOS

Engineering of a Nepetalactol-Producing Platform Strain of Saccharomyces cerevisiae for the Production of Plant Seco-Iridoids

ACS Synthetic biology

Campbell, Alex; Bauchart, Philippe; Gold, Nicholas D.; Zhu, Yun; De Luca, Vincenzo; Martin, Vincent J. J.

<P>The monoterpene indole alkaloids (MIAs) are a valuable family of chemicals that include the anticancer drugs vinblastine and vincristine. These compounds are of global significance&#58872;appearing on the World Health Organization&rsquo;s list of model essential medicines&#58872;but remain exorbitantly priced due to low <I>in planta</I> levels. Chemical synthesis and genetic manipulation of MIA producing plants such as <I><I>Catharanthus roseus</I></I> have so far failed to find a solution to this problem. Synthetic biology holds a potential answer, by building the pathway into more tractable organisms such as <I><I>Saccharomyces cerevisiae</I></I>. Recent work has taken the first steps in this direction by producing small amounts of the intermediate strictosidine in yeast. In order to help improve on these titers, we aimed to optimize the early biosynthetic steps of the MIA pathway to the metabolite nepetalactol. We combined a number of strategies to create a base strain producing 11.4 mg/L of the precursor geraniol. We also show production of the critical intermediate 10-hydroxygeraniol and demonstrate nepetalactol production <I>in vitro</I>. Lastly we demonstrate that activity of the iridoid synthase toward the intermediates geraniol and 10-hydroxygeraniol results in the synthesis of the nonproductive intermediates citronellol and 10-hydroxycitronellol. This discovery has serious implications for the reconstruction of the MIA in heterologous organisms.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/asbcd6/2016/asbcd6.2016.5.issue-5/acssynbio.5b00289/production/images/medium/sb-2015-00289f_0007.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/sb5b00289'>ACS Electronic Supporting Info</A></P>

2016

American Chemical Society

cc-by-nc

2161-5063

5

5

pp.405-414

10.1021/acssynbio.5b00289.s001

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART75509046

https://figshare.com/articles/journal_contribution/Engineering_of_a_Nepetalactol_Producing_Platform_Strain_of_i_Saccharomyces_cerevisiae_i_for_the_Production_of_Plant_Seco_Iridoids/3124555/1/files/4861681.pdf

monoterpene indole alkaloids; seco-iridoid; Saccharomyces cerevisiae; 10-hydroxygeraniol; nepetalactol; iridodial