ATP sulfurylase (ATPS) catalyzes the first committed step in the sulfate assimilation pathway, the activation of sulfate prior to its reduction. ATPS has been studied in only a few model organisms and even in these cases to a much smaller extent than the sulfate reduction and cysteine synthesis enzymes. This is possibly because the latter were considered of greater regulatory importance for sulfate assimilation. Recent evidences (reported in this paper) challenge this view and suggest that ATPS may have a crucial regulatory role in sulfate assimilation, at least in algae. In the ensuing text, we summarize the current knowledge on ATPS, with special attention to the processes that control its activity and gene(s) expression in algae. Special attention is given to algae ATPS proteins. The focus on algae is the consequence of the fact that a comprehensive investigation of ATPS revealed that the algal enzymes, especially those that are most likely involved in the pathway of sulfate reduction to cysteine, possess features that are not present in other organisms. Remarkably, algal ATPS proteins show a great diversity of isoforms and a high content of cysteine residues, whose positions are often conserved. According to the occurrence of cysteine residues, the ATPS of eukaryotic algae is closer to that of marine cyanobacteria of the genera Synechococcus and Prochlorococcus and is more distant from that of freshwater cyanobacteria. These characteristics might have evolved in parallel with the radiation of algae in the oceans and the increase of sulfate concentration in seawater.
The target of rapamycin (TOR) signaling pathway regulates fundamental intracellular functions critical for cell viability and proliferation. Manipulation of TOR in high lipid-producing microalgae may help overcome the trade-off between biomass production and lipid yield that still impairs the viable production of biofuel from microalgae. In this study, we inhibited the TOR kinase in the model diatom Phaeodactylum tricornutum using the selective TOR inhibitor AZD-8055, and analyzed cell proliferation, chlorophyll content, lipid synthesis and carbon metabolism. AZD-8055 inhibits cell proliferation in a dose-dependent manner compared to N deprivation which stops growth. Microscopy, flow cytometry, and quantitative analyses of lipids also demonstrated that AZD-8055 treatment strongly promotes triacylglycerol (TAG) accumulation while decreasing the quantity of sterols. The TAG productivity of AZD-8055 treated cultures was significantly higher than for N deprived cultures. Measurement of the activities of the key metabolic enzymes glyceraldehyde phosphate dehydrogenase (GAPDH), glucose-6-phosphate dehydrogenase (G6PDH) and malate dehydrogenase (MDH) revealed opposite effects for AZD-8055 treatment and N-starvation on the activity of the glycolytic enzyme GAPDH. This suggests that TOR inhibition and N starvation may have distinct impacts on general metabolism and lipid accumulation. Our main finding is that treating cultures with AZD-8055 results in higher TAG productivity than N starvation in P. tricornutum. The chemical or genetic manipulation of the TOR signaling pathway in P. tricornutum and other diatoms may lead to the development of strains or approaches suitable for the enhanced production of TAGs for biofuel
The presence of CO2-concentrating mechanisms (CCMs) is believed to be one of the characteristics that allows diatoms to thrive in many environments and to be major contributors to global productivity. Here, the type of CCM and the responses to variable CO2 concentration were studied in marine and freshwater diatoms. At 400 ppm, there was a large diversity in physiological and biochemical mechanisms among the species. While Phaeodactylum tricornutum mainly used HCO3-, Thalassiosira pseudonana mainly used CO2. Carbonic anhydrase was an important component of the CCM in all species and C4 metabolism was absent, even with T. weissflogii. For all species, at 20 000 ppm, the affinity for dissolved inorganic carbon was lower than at 400 ppm CO2 and the reliance on CO2 was higher. Despite the difference in availability of inorganic carbon in marine and fresh waters, there were only small differences in CCMs between species from the two environments, and Navicula pelliculosa behaved similarly when grown in the two environments. The results suggest that species-specific differences are great, and more important than environmental differences in determining the nature and effectiveness of the CCM in diatoms.
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