Two stralns of the paralytic shellfish tox~n (PST) producing dinoflagellate Alexandrium lninutum Halim (highly toxic ALlV and weakly toxic AL2V) were grown in batch culture with either nitrate or phosphate as the limiting nutrient. In comparison with cells of the strain ALlV, cells of AL2V grew at a similar C-specific rate, had a higher C/N ratio, and lower ratios of chl a/chl c2 and chl a/peridinin. Neither chlorophylls nor carotenoids could be used to estimate C-biomass, N-biomass or toxin content for this organism. The toxin profile for both strains was dominated (up to 95 %) by the gonyautoxin GTX4, with smaller proportions of GTX1. GTX2 and GTX3. The rate of toxin synthesis for both strains was greatest 1 to 2 d after the N-refeeding of N-deprived cells, with the net rate of toxin synthesis exceeding that of C-biomass and cell division by a factor of up to 4. Toxin synthesis was not enhanced by short-term P-stress. N-stress alone led to a decrease in toxin cell-', but P-stress followed by N-stress did not result in such a decline, implicating phosphorus in the regulation of toxin metabolism. Although arginine is a major precursor for PST synthesis, taurine, glycine, glutamine, and cell N showed similar relations to that observed for arginine with respect to toxin content. Furthermore, the mole ratio of arginine/toxin could vary by a factor of up to 5 between ALlV and AL2V at peak values of toxin cell-', and by more than 5 within a strain when growing under different conditions. These observations suggest that the relationship between free arginine content and toxin content is complex. No explanation for the higher toxin content of A L l V IS apparent, except that ALlV has a higher N-content per cell and this may be conducive to a higher rate of synthesis of the N-rich toxins.

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Oxygen-balanced mixotrophy (OBM) is a novel type of microalgal cultivation that improves autotrophic productivity while reducing aeration costs and achieving high biomass yields on substrate. The scale-up of this process is not straightforward, as nonideal mixing in large photobioreactors might have unwanted effects in cell physiology. We simulated at lab scale dissolved oxygen and glucose fluctuations in a tubular photobioreactor operated under OBM where glucose is injected at the beginning of the tubular section. We ran repeated batch experiments with the strain Galdieria sulphuraria ACUF 064 under glucose pulse feeding of different lengths, representing different retention times: 112, 71, and 21 min. During the long and medium tube retention time simulations, dissolved oxygen was depleted 15-25 min after every glucose pulse. These periods of oxygen limitation resulted in the accumulation of coproporphyrin III in the supernatant, an indication of disruption in the chlorophyll synthesis pathway. Accordingly, the absorption cross-section of the cultures decreased steeply, going from values of 150-180 m 2 kg −1 at the end of the first batch down to 50-70 m 2 kg −1 in the last batches of both conditions. In the short tube retention time simulation, dissolved oxygen always stayed above 10% air saturation and no pigment reduction nor coproporphyrin III accumulation were observed. Concerning glucose utilization efficiency, glucose pulse feeding caused a reduction of biomass yield on substrate in the range of 4%-22% compared to the maximum levels previously obtained with continuous glucose feeding (0.9 C-g C-g −1 ).The missing carbon was excreted to the supernatant as extracellular polymeric substances constituted by carbohydrates and proteins. Overall, the results point out the importance of studying large-scale conditions in a controlled environment and the need for a highly controlled glucose feeding strategy in the scale-up of mixotrophic cultivation.