Dual-plasmid interactions stimulate the accumulation of valencene in Saccharomyces cerevisiae

Plasmids are one of the most commonly used basic tools in the construction of microbial cell factories, the use of which individually or in pairs play an important role in the expression of exogenous gene modules. However, little attention has been paid to the interactions of plasmid-plasmid and plasmid-host in the widespread use of the double plasmid system. In this study, we demonstrated that dual-plasmid interactions facilitated to cell growth and product accumulation in Saccharomyces cerevisiae. The strain containing both the expression plasmid pEV (a plasmid carrying the gene encoding valencene synthase) and the assistant plasmid pI (an empty plasmid expressing no extra gene) showed a significant improvement in relative growth rate, biomass and valencene production compared to the strain containing only the pEV plasmid. The transcriptional level analysis revealed an up-regulated expression of specific gene on the expression plasmid pEV stimulated by the assistant plasmid pI in the dual-plasmid interactions. Further investigations demonstrated the essential roles of the promoters of the expression plasmid pEV and the CEN/ARS element of the assistant plasmid pI in the dual-plasmid interactions. Combined with the results of predicted nucleosome occupancy, a response model of interaction based on the key T(n)C and CEN/ARS element was established. Moreover, the transformation order of the two plasmids significantly affected the response effect, implying the dominance of plasmid pI in the dual-plasmid interactions. Our finding first demonstrated that dual plasmids regulate the gene expression through spatial interactions at DNA sequences level, which provides a new perspective for the development of microbial cell factories in future.