Identification of Novel Knockout Targets for Improving Terpenoids Biosynthesis in Saccharomyces cerevisiae

Abstract

Many terpenoids have important pharmacological activity and commercial value; however, application of these terpenoids is often limited by problems associated with the production of sufficient amounts of these molecules. The use of Saccharomyces cerevisiae (S. cerevisiae) for the production of heterologous terpenoids has achieved some success. The objective of this study was to identify S. cerevisiae knockout targets for improving the synthesis of heterologous terpeniods. On the basis of computational analysis of the S. cerevisiae metabolic network, we identified the knockout sites with the potential to promote terpenoid production and the corresponding single mutant was constructed by molecular manipulations. The growth rates of these strains were measured and the results indicated that the gene deletion had no adverse effects. Using the expression of amorphadiene biosynthesis as a testing model, the gene deletion was assessed for its effect on the production of exogenous terpenoids. The results showed that the dysfunction of most genes led to increased production of amorphadiene. The yield of amorphadiene produced by most single mutants was 8–10-fold greater compared to the wild type, indicating that the knockout sites can be engineered to promote the synthesis of exogenous terpenoids.

Heterologous biosynthesis of artemisinic acid in Saccharomyces cerevisiae

“…Sun et al . () predicted some knockout sites that can improve the titre of terpenoids but not inhibit cell growth in silico and in vivo . Another attempt identifies the knockout sites for increasing dihydroartemisinic acid production based on in silico FBA and minimization of metabolic adjustment on a genome‐scale model (Misra et al .…”

“…As mentioned above, the traditional strategies result in dysfunction of metabolism (Sun et al 2014). To address this issue, iMM904, a genome-scale metabolic model of S. cerevisiae has been developed (Zomorrodi and Maranas 2010).…”

“…Combining with flux balance analysis (FBA) and minimization of metabolic adjustment, the knockout sites can be predicted. Sun et al (2014) predicted some knockout sites that can improve the titre of terpenoids but not inhibit cell growth in silico and in vivo.…”

“…However, the simple up‐regulation or down‐regulation of key genes may result in dysfunction of metabolism, subsequently, the growth of the cell could be inhibited by the accumulation of toxic intermediates (Sun et al . ).…”

“…All these traditional strategies mainly focused on increasing carbon flux towards heterologous biosynthesis pathway in host cells. However, the simple up-regulation or down-regulation of key genes may result in dysfunction of metabolism, subsequently, the growth of the cell could be inhibited by the accumulation of toxic intermediates (Sun et al 2014).…”