Advancing Yarrowia lipolytica sub-organelle engineering with endogenous mitochondrial targeting sequence.

OBJECTIVE

The aim of the study was identification and validation of an endogenous mitochondrial targeting signal (MTS) sequence of Yarrowia lipolytica, for efficient compartmentalization of a target protein to mitochondria.

RESULTS

MTS from citrate synthase of Y. lipolytica (YlCISY-MTS) was identified, isolated and fused with green fluorescent protein (GFP) to direct it to the mitochondrial matrix. The efficiency of localization of GFP to mitochondrial matrix with YlCISY-MTS was compared with currently used MTS from Saccharomyces cerevisiae's cytochrome oxidase subunit IV. Confocal microscopy confirmed the targeted and greater GFP localization, underlining the potential of endogenous YlCISY-MTS for mitochondrial engineering in Y. lipolytica. The availability of endogenous MTS will evade the need of codon optimization of S. cerevisiae MTS for mitochondrial engineering in Y. lipolytica. This is the first report of an endogenous MTS of Y. lipolytica.

CONCLUSION

An endogenous MTS of Y. lipolytica has been identified to facilitate the targeted delivery of a protein in the mitochondria enabling future advancements through leveraging the unique subcellular environment for metabolic engineering applications.