Abstract
Oleaginous yeast Rhodosporidium toruloides has great biotechnological potential and scientific interest, yet the molecular rationale of its cellular behavior to carbon and nitrogen ratios with concurrent lipid agglomeration remains elusive. Here, metabolomics adaptations of the R. toruloides in response to varying glucose and nitrogen concentrations have been investigated. In preliminary screening we found that 5% glucose (w/v) was optimal for further analysis in Rhodosporidium toruloides 3641. Hereafter, the effect of complementation to increase lipid agglomeration was evaluated with different nitrogen sources and their concentration. The results obtained illustrated that the biomass (13 g/L) and lipid (9.1 g/L) production were maximum on 5% (w/v) glucose and 0.12% (NH4)2SO4. Furthermore, to shed lights on lipid accumulation induced by nitrogen-limitation, we performed metabolomic analysis of the oleaginous yeast R. toruloides 3641. Significant changes were observed in metabolite concentrations by qualitative metabolomics through gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), which were mapped onto the governing metabolic pathways. Notable finding in this strain concerns glycerol and CDP-DAG metabolism wherein reduced production of glycerol and phospholipids induced a bypass leading to enhanced de-novo triacylglyceride synthesis. Collectively, our findings help in understanding the central carbon metabolism of R. toruloides which may assist in developing rationale metabolic models and engineering efforts in this organism.
Highlights
IntroductionThe red yeast Rhodosporidium toruloides has been recognized as a promising microbial cell factory for the production of functional lipids, oleo-chemicals, and biofuels [1,2]
Several reports concerning the lipid agglomeration of R. toruloides have been implemented using (NH4 )2 SO4 as the nitrogen source; the metabolomics study and glycerol production during the cultivation has not been addressed
Several reports concerning the lipid agglomeration of R. toruloides have been implemented using (NH4)2SO4 as the nitrogen source; the metabolomics study and glycerol production during the cultivation has not been ad5 of 20 dressed
Summary
The red yeast Rhodosporidium toruloides has been recognized as a promising microbial cell factory for the production of functional lipids, oleo-chemicals, and biofuels [1,2]. Triacylglycerols (TAG) is the major form of neutral lipids (up to 70% of dry cell weight) accumulated in this yeast. This basidiomycetous fungus is a potential host for metabolic engineering to produce terpenes and fatty acids [3]. R. toruloides can metabolize challenging substrates including waste glycerol, biomass hydrolysates, xylose, and can naturally co-produce valuable compounds such as carotenoids and other useful enzymes relevant to the pharmaceutical and chemical industries [2,3,4]
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