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This study aims to elucidate the role of light absorption rate and nitrate concentration on triacylglycerol (TAG) productivity and cellular TAG accumulation in nitrogen limited Parachlorella kessleri cultures grown in chemostat mode. In batch operated nitrogen starved cultures the cells accumulate large amounts of TAGs at the expense of biomass growth leading to severe decrease in cell division rate prompting eventual cell death. In PBRs operated in continuous mode cell multiplication can persist in nitrogen limited conditions with simultaneous TAG accumulation. P. kessleri cultures were grown in a flat-plate PBR with constant dilution rate of 0.01 1/h and feed-medium nitrate concentration ranging from 1 to 16 mM. The steady-state biomass, pigment, carbohydrate, protein, total lipid, and TAG concentrations as well as the spectral absorption and scattering cross-sections of the microalgae were measured for each culture. These were used to estimate the specific mean rate of photon absorption (MRPA) in the PBR. Maximum areal TAG productivity of 2.6 g/m 2 ⋅day was obtained for the culture grown with nitrate concentration equal to 3.65 mM and a mean rate of photon absorption equal to 17,700 μmol hν /kg X A s. TAG productivity and cellular TAG accumulation varied with the nitrate concentration in the feed medium and MRPA. Finally, our results show that TAG productivity can be further increased by optimizing the mean rate of photon absorption through adjusting culture dilution rate and feed-medium nitrate concentration.

The pharmaceutical industry faces a growing demand and recurrent shortages in many anticancer plant drugs given their extensive use in human chemotherapy. Efficient alternative strategies of supply of these natural products such as bioproduction by microorganisms are needed to ensure stable and massive manufacturing. Here, we developed and optimized yeast cell factories efficiently converting tabersonine to vindoline, a precursor of the major anticancer alkaloids vinblastine and vincristine. First, fine-tuning of heterologous gene copies restrained side metabolites synthesis towards vindoline production. Tabersonine to vindoline bioconversion was further enhanced through a rational medium optimization (pH, composition) and a sequential feeding strategy. Finally, a vindoline titre of 266 mg l À1 (88% yield) was reached in an optimized fed-batch bioreactor. This precursordirected synthesis of vindoline thus paves the way towards future industrial bioproduction through the valorization of abundant tabersonine resources.

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Energy-storage metabolites such as neutral lipids and carbohydrates are valuable compounds for liquid biofuel production. The aim of this work is to elucidate the main biological responses of two algae species known for their effective energy-rich compound accumulation in nitrogen limitation and day–night cycles: Nannochloropsis gaditana, a seawater species, and Parachlorella kessleri, a freshwater species. Lipid and carbohydrate production are investigated, as well as cell resistance to mechanical disruption for energy-rich compound release. Nitrogen-depleted N. gaditana showed only a low consumption of energy-storage molecules with a non-significant preference for neutral lipids (TAG) and carbohydrates in day–night cycles. However, it did accumulate significantly fewer carbohydrates than P. kessleri. Following this, the highest levels of productivity for N. gaditana in chemostat cultures at four levels of nitrogen limitation were found to be 3.4 and 2.2 ×10−3 kg/m2·d for carbohydrates and TAG, respectively, at 56%NO3 limitation. The cell disruption rate of N. gaditana decreased along with nitrogen limitation, from 75% (at 200%NO3) to 17% (at 13%NO3). In the context of potentially recoverable energy for biofuels, P. kessleri showed good potential for biodiesel and high potential for bioethanol; by contrast, N. gaditana was found to be more efficient for biodiesel production only.

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Nitrogen limitation and changing solar conditions are both known to affect triacylglycerol (TAG) production in microalgae. This study investigates the optimization of TAG production with a continuous nitrogen-limited culture of Nannochloropsis gaditana in simulated day-night cycles (DNc). The effect of DNc was first investigated in nitrogen-deprived condition (i.e., batch culture), emphasizing a significant change in mechanical resistance of the strain during the night. The