Metabolomic Profiling Revealed Diversion of Cytidinediphosphate-Diacylglycerol and Glycerol Pathway towards Denovo Triacylglycerol Synthesis in Rhodosporidium toruloides
Abstract
:1. Introduction
2. Materials and Methods
2.1. Yeast and Culture Conditions
2.2. Biochemical Analysis
2.3. Lipid Quantification and Profiling
2.4. Lipid Droplet Size Estimation
2.5. Qualitative Metabolomics Using GC-MS
2.6. LC–MS Based Untargeted Metabolite Analysis
2.7. Statistical Analysis
3. Results
3.1. Cultivation of R. toruloides at Variable Carbon Concentrations
3.2. Time Course of Lipid Accumulation under Nitrogen-Limited Condition
3.3. R. toruloideslipid Droplet Formation at Nitrogen Stress Condition
3.4. Fatty Acid Profiling of R. toruloides under Nutrient Stress Condition
3.5. GC–MS and LC-MS/MS Based Untargeted Metabolome Analysis
3.5.1. GC-MS Analysis
3.5.2. LC-MS/MS Analysis
3.6. Metabolic Pathway Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
Abbreviations
References
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YNB Medium | DCW (g/L) | Lipid Titer (g/L) | Glycerol (g/L) | Citric Acid (g/L) | Residual Glucose (g/L) | Butanediol (g/L) | FAME Productivity (g/L/h) |
---|---|---|---|---|---|---|---|
3% Glucose, 0.5% (NH4)2SO4 | 6.8 ± 0.18 | 3.4 ± 0.08 | - | - | 1.67 ± 0.005 | - | 0.53 ± 0.02 |
4% Glucose, 0.5% (NH4)2SO4 | 8.2 ± 0.15 | 4 ± 0.007 | 3.45 ± 0.038 | 0.52 ± 0.005 | 2.58 ± 0.006 | 0.11 ±0.005 | 0.48 ± 0.04 |
5% Glucose, 0.5% (NH4)2SO4 | 9.6 ± 0.12 | 5.8 ± 0.08 | 3.83 ± 0.027 | 0.77 ± 0.008 | 4.26 ± 0.007 | 0.12 ± 0.006 | 0.69 ± 0.02 |
6% Glucose, 0.5% (NH4)2SO4 | 10.4 ± 0.28 | 5.3 ± 0.12 | 6.27 ± 0.045 | 0.43 ± 0.007 | 10.29 ± 0.004 | 0.07 ± 0.008 | 0.63 ± 0.05 |
7% Glucose, 0.5% (NH4)2SO4 | 9.2 ± 0.13 | 3.8 ± 0.15 | 5.60 ± 0.036 | 0.49 ± 0.006 | 21.28 ± 0.016 | - | 0.45 ± 0.03 |
8% Glucose, 0.5% (NH4)2SO4 | 9.2 ± 0.27 | 3.6 ± 0.08 | 6.63 ± 0.042 | 0.48 ± 0.008 | 35.03 ± 0.015 | 0.07 ± 0.008 | 0.43 ± 0.03 |
9% Glucose, 0.5% (NH4)2SO4 | 7.41 ± 0.19 | 3.1 ± 0.06 | 3.84 ± 0.054 | 0.67 ± 0.009 | 36.89 ± 0.008 | 0.05 ± 0.004 | 0.37 ± 0.05 |
10% Glucose, 0.5% (NH4)2SO4 | 7.25 ± 0.17 | 2.5 ± 0.04 | 3.67 ± 0.044 | 0.70 ± 0.045 | 44.98 ± 0.014 | 0.08 ± 0.006 | 0.3 ± 0.04 |
5% Glucose, 0.5% Yeast extract | 10.2 ± 0.28 | 3.1 ± 0.16 | 6.34 ± 0.45 | 1.10 ± 0.52 | 8.75 ± 0.48 | - | 0.37 ± 0.03 |
5% Glucose, 0.5% Peptone | 8.1 ± 0.27 | 3.6 ± 0.18 | 2.51 ± 0.34 | 1.02 ± 0.37 | 2.60 ± 0.35 | 0.12 ± 0.06 | 0.43 ± 0.06 |
5% Glucose, 0.5% Urea | 5.5 ± 0.14 | 2.8 ± 0.16 | - | - | 10.68 ± 0.57 | - | 0.33 ± 0.04 |
5% Glucose, 0.12% (NH4)2SO4, | 13 ± 0.52 | 8.23 ± 0.48 | 1.72 ± 0.08 | 0.08 ± 0.004 | 5.2 ± 0.26 | 0.13 ± 0.05 | 0.98 ± 0.06 |
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Deeba, F.; Kumar, K.K.; Rajacharya, G.H.; Gaur, N.A. Metabolomic Profiling Revealed Diversion of Cytidinediphosphate-Diacylglycerol and Glycerol Pathway towards Denovo Triacylglycerol Synthesis in Rhodosporidium toruloides. J. Fungi 2021, 7, 967. https://doi.org/10.3390/jof7110967
Deeba F, Kumar KK, Rajacharya GH, Gaur NA. Metabolomic Profiling Revealed Diversion of Cytidinediphosphate-Diacylglycerol and Glycerol Pathway towards Denovo Triacylglycerol Synthesis in Rhodosporidium toruloides. Journal of Fungi. 2021; 7(11):967. https://doi.org/10.3390/jof7110967
Chicago/Turabian StyleDeeba, Farha, Kukkala Kiran Kumar, Girish H. Rajacharya, and Naseem A. Gaur. 2021. "Metabolomic Profiling Revealed Diversion of Cytidinediphosphate-Diacylglycerol and Glycerol Pathway towards Denovo Triacylglycerol Synthesis in Rhodosporidium toruloides" Journal of Fungi 7, no. 11: 967. https://doi.org/10.3390/jof7110967