Transcriptomic and metabolomic profiling of a Rhodotorula color mutant to improve its lipid productivity in fed-batch...

Lipids produced by oleaginous microorganisms enrich the supply of feedstock for bio-fuel. In this study, a mutant (Mut) obtained by UV-nitrosoguanidine mutagenesis turned its colony color from orange-red to light-yellow and showed higher lipid productivity at 37 °C than the original strain Rhodotorula sp. U13N3 (Rht) in the glycerol medium. The metabolic changes between Mut and Rht in batch fermentation were investigated by transcriptomic and metabolomic profiling at the biomass accumulation (30 h) and lipid production (96 h) stages. The average base number in each strain was 5.80 × 10⁹ ± 0.38 × 10⁹ bp (mean ± SD) with 62.43% ± 0.13% GC ratio, and 7499 unigenes were assembled after Illumina sequencing. Moreover, 33 metabolites were quantified by ¹H NMR-based profiling. The multi-omics results demonstrated that Mut showed increased glycerol transport and utilization capabilities especially at the first stage (30 h). Then the carbon flux shifted from the TCA cycle to lipid production (96 h). The increased lipid productivity of Mut was partially attributed to the down-regulation of mannitol biosynthesis. However, the mechanism for color change was elusive. At 96 h, the low level of cytosol glycerol probably restricted the lipid production. As a result, supplementation of glycerol in fed-batch fermentation remarkably improved the biomass, lipid production, and lipid content to 34.60 g/L, 25.72 g/L, and 74.3% (w/w dcw), respectively. The cell morphology implied that excessively prolonging the fermentation time was detrimental to the final lipid yield due to cell breakage. In conclusion, the Rhodotorula mutant provided a candidate strain for lipid production with glycerol as the carbon source.