Characterization of the endogenous promoters in Yarrowia lipolytica for the biomanufacturing applications

Abstract

Abstract Background: Yarrowia lipolytica, an oleaginous yeast with the GRAS status, has been developed as a platform for the chemical production. Specifically, promoter engineering is an important approach to regulating gene expression at the transcriptional level, which is of great significance in constructing microbial cell factories. Until now, some work on the promoter study has been carried out in Y. lipolytica. However, compared with other microorganisms, such as Saccharomyces cerevisiae, Bacillus subtilis, and etc., obtained achievements still need further replenishment and development for Y. lipolytica. Results: To accurately analyze the promoter strength and avoid background interference, we used the NanoLuc luciferase reporter method. Furthermore, we screened 81 endogenous promoters in Y. lipolytica, mainly involved in carbon metabolism, amino acid metabolism, and lipid metabolism. Among them, the strongest promoter is PMnDH2 (YALI0D18964g), 1.60-fold of the strength of the PTEF promoter, and the weakest promoter is PPHO89 (YALI0E23859g), 0.06% of the PTEF promoter. Conclusions: As a result, we obtained 15 strong promoters, 41 medium strength promoters, and 25 weak promoters, with the strength spanning from 0.06% to 1.60-fold of PTEF promoter. In general, our study provides a unique and available endogenous promoter library for Y. lipolytica.