| dc.description.abstract | Rhodotorula toruloides is an oleaginous yeast with potential use as a biotechnological chassis for both production of industrially and pharmaceutically relevant compounds, and as a drop-in biofuel producer on low-cost substrate. Cells can accumulate lipid droplets to over 70 % weight/weight under certain growth conditions. We here summarise the currently-available genetic and molecular toolkit for the yeast and suggest further avenues for research to enable full utilisation of this yeast. To aid in these objectives, we have constructed lipid droplet-associated GFP-tagged protein Ldp1-GFP and demonstrated how this can be used to quantify individual cell lipid quantity using confocal microscopy. Calnexin-GFP and GFP-Atg8 have also been constructed for live-cell monitoring of the intracellular machinery in lipid droplet synthesis as part of the developing R. toruloides molecular toolkit. Furthermore, the induction profiles of selected heat shock protein promoters have been characterised through a GFP-reporter system. Currently, the only reported inducible promoters in R. toruloides are nutrient-dependent NAR1, ICL1, MET16, CTR3, DAO1, THI4, THI5 and CTR31, and tightly controlling these is not possible in potential use as a biofuel producer using low-cost substrate. Of the promoters highlighted herein (ENO2, TDH3, PGK1, TPI1, SSB1, ACT1 and TDH3), ENO2 is identified as a qualitatively putative heat-shock inducible promoter, further analysis of which may allow the nutrient-dependency limitation to be overcome through exploitation of the native environmental stress response. | en_GB |
