Enhancement of the lipid productivity and fatty acid methyl ester profile of Chlorella vulgaris by two rounds of mutagenesis

In this study, we applied a second round of random mutagenesis using ethyl methanesulfonate to further increase the lipid productivity of a Chlorella vulgaris mutant strain. We generated a mutant (UV715-EMS25) with a lipid content and biomass that were respectively 67% and 35% higher than those of the wild type (WT). The highest achieved lipid productivity in UV715-EMS25 was 91 mg L-1 day(-1). Gas chromatography-mass spectrophotometric analysis revealed that the fatty acid methyl ester content of the mutant was 3.9-fold higher compared with that of WT cells. Amounts of saturated and monounsaturated fatty acids were also higher in the mutant, while the total amounts of polyunsaturated fatty acids were lower. Finally, the mutant displayed superior lipid productivity compared with the WT during pilot-scale cultivation in a flat panel photobioreactor. All these results demonstrate that UV715-EMS25 is highly suitable for biodiesel production

Understanding lipid metabolism in high-lipid-producing Chlorella vulgaris mutants at the genome-wide level

In this study, physical and chemical mutagenesis methods were applied to enhance lipid productivity in Chlorella vulgaris. Then, de novo RNA-seq was performed to observe lipid metabolism changes at the genome-wide level. Characterization of two mutants, UV-715 and EMS-25, showed marked increases in lipid contents, i.e., 42% and 45%, respectively. In addition, the biomass productivity of the UV-715 cells was 9% higher than that of wildtype cells. Furthermore, gas chromatography-mass spectrophotometry analysis showed that both mutants have higher fatty acid methyl ester (FAME) contents than wild-type cells. To understand the effect of mutations that caused yield changes in UV-715 and EMS-25 cells at a genome-wide level, we carried out de novo RNA-seq. As expected, the transcriptional levels of the lipid biosynthesis genes were up-regulated, while the transcriptional levels of genes involved in lipid catabolism were down-regulated. Surprisingly, the transcriptional levels of the genes involved in nitrate assimilation and detoxification of reactive oxygen species (ROS) were significantly increased in the mutants. The genome-wide analysis results highlight the importance of nitrate metabolism and detoxification of ROS for high biomass and lipid productivity

Transcriptomic and fatty acid analyses of Neochloris aquatica grown under different nitrogen concentration

In this study, we characterized the fatty acid production in Neochloris aquatica at transcriptomics and biochemical levels under limiting, normal, and excess nitrate concentrations in different growth phases. At the stationary phase, N. aquatica mainly produced saturated fatty acids such as stearic acid under the limiting nitrate concentration, which is suitable for biodiesel production. However, it produced polyunsaturated fatty acids such as alpha-linolenic acid under the excess nitrate concentration, which has nutritional values as food supplements. In addition, RNA-seq was employed to identify genes and pathways that were being affected in N. aquatica for three growth phases in the presence of the different nitrate amounts. Genes that are responsible for the production of saturated fatty acids were upregulated in the cells grown under a limiting nitrogen amount while genes that are responsible for the production of polyunsaturated fatty acid were upregulated in the cells grown under excess nitrogen amount. Further analysis showed more genes differentially expressed (DEGs) at the logarithmic phase in all conditions while a relatively steady trend was observed during the transition from the logarithmic phase to the stationary phase under limiting and excess nitrogen. Our results provide a foundation for identifying developmentally important genes and understanding the biological processes in the different growth phases of the N. aquatica in terms of biomass and lipid production