No abstract

Microalgae are a promising feedstock for biofuel production. Microalgal metabolic pathways are heavily influenced by environmental factors. For instance, lipid metabolism can be induced by nitrogen-limiting conditions. However, the underlying mechanisms of lipid biosynthesis are unclear. In this study, we analyzed the global metabolic profiles of three genetically closely related Chlorella strains (C1, C2, and C3) with significant differences in lipid productivity to identify the contributions of key metabolic pathways to lipid metabolism. We found that nitrogen obtained from amino acid catabolism was assimilated via the glutamate–glutamine pathway and then stored as amino acids and intermediate molecules (particularly proline, alanine, arginine, succinate, and gamma-aminobutyrate) via the corresponding metabolic pathways, which led to carbon–nitrogen disequilibrium. Excess carbon obtained from photosynthesis or glycolysis was re-distributed into carbon-containing compounds, such as glucose-6-phosphate, fructose-6-phosphate, phosphoenolpyruvate, lactate, citrate, 3-hydroxybutyrate, and leucine, and then diverted into lipid metabolism for the production of storage lipids via the gamma-aminobutyrate pathway, glycolysis, and the tricarboxylic acid cycle. These results were substantiated in the model green alga Chlamydomonas reinhardtii by analyzing various mutants deficient in glutamate synthase/NADH-dependent, glutamate synthase/Fd-dependent, glutamine synthetase, aspartate aminotransferase, alanine aminotransferase, pyruvate kinase, and citrate synthase. Our study suggests that not only carbon but also nitrogen assimilation and distribution pathways contribute to lipid biosynthesis. Furthermore, these findings may facilitate genetic engineering efforts to enhance microalgal biofuel production.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0839-4) contains supplementary material, which is available to authorized users.

Nitrogen is an essential nutrient element. Ammonium nitrogen, one of the most common nitrogen sources, is found in various habitats, especially wastewater. However, excessive amounts of ammonium nitrogen can be toxic to phytoplankton, higher plants, fish, and other animals, and microorganisms. In this study, we explored the tolerance of green algae to ammonium nitrogen using 10 Chlorella strains. High concentrations of ammonium nitrogen directly inhibited the growth of Chlorella, but the degree of inhibition varied by strain. With the EC50 of 1.6 and 0.4 g L−1, FACHB-1563 and FACHB-1216, respectively had the highest and lowest tolerance to ammonium nitrogen among all strains tested, suggesting that FACHB-1563 could potentially be used to remove excess ammonium nitrogen from wastewater in bioremediation efforts. Two strains with the highest and lowest tolerance to ammonium nitrogen were selected to further explore the inhibitory effect of ammonium nitrogen on Chlorella. Analysis of chlorophyll fluorescence, oxygen evolution, and photosynthesis proteins via immunoblot showed that photosystem II (PSII) had been damaged when exposed to high levels of ammonium nitrogen, with the oxygen-evolving complex as the primary site, and electron transport from QA− to QB was subsequently inhibited by this treatment. A working model of ammonium nitrogen competition between N assimilation and PSII damage is proposed to elucidate that the assimilation rate of ammonium nitrogen by algae strains determines the tolerance of cells to ammonium nitrogen toxicity.