Mechanism and enhancement of lipid accumulation in filamentous oleaginous microalgae Tribonema minus under heterotrophic condition

BACKGROUND: The filamentous microalgae Tribonema minus accumulates large amounts of lipids under photoautotrophic condition, while under heterotrophic condition, the lipid content decreased dramatically. Determination of the differences in metabolic pathways between photoautotrophic and heterotrophic growth will provide targets and strategies for improvement of lipid accumulation in heterotrophic cells. METHODS: The metabolic differences between photoautotrophically and heterotrophically cultivated T. minus cells were studied by comparing the growth, biochemical compositions and transcriptomic and metabolomic profiles of the cells. Based on comparative transcriptomic and metabolomic studies, we generated a global model of the changes in central carbon metabolism and lipid biosynthetic pathways that occur under photoautotrophic and heterotrophic growth conditions. Moreover, the specific effects of supplementation with exogenous key metabolic intermediates on the lipid accumulation in heterotrophic culture were analyzed. RESULTS: Compared to photoautotrophic cultures, heterotrophic cultures exhibited enhanced biomass levels and carbohydrate content, but decreased lipid accumulation. These effects were accompanied by low expression levels of genes involved in glycolysis, de novo fatty acids biosynthesis and lipid biosynthesis, and high levels of genes involved in gluconeogenesis. In addition, the levels of key metabolites involved in glycolysis/gluconeogenesis were elevated in abundance, whereas those of certain fatty acids and citric acid were decreased in heterotrophic cultures. Upon supplementation with exogenous potassium palmitate, the lipid content increased dramatically in heterotrophically cultivated T. minus. CONCLUSION: An insufficient supply of carbon precursors caused the low levels of lipid accumulation during heterotrophic cultivation. Appropriate carbon metabolite supplementation based on the metabolomic data was shown to promote lipid accumulation. Moreover, gene regulatory metabolic targets were also identified via omics analysis.