Marine microalgae have received much attention as a sustainable source of the two health beneficial omega-3-fatty acids docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5). However, photoautotrophic monocultures of microalgae can only produce either DHA or EPA enriched biomass. An alternative may be the photoautotrophic co-cultivation of Tisochrysis lutea as DHA-producer with Microchloropsis salina for simultaneous EPA production to obtain EPA- and DHA-rich microalgae biomass in a nutritionally balanced ratio. Photoautotrophic co-cultivation processes of T. lutea and M. salina were studied, applying scalable and fully controlled lab-scale gas-lift flat-plate photobioreactors with LED illumination for dynamic climate simulation of a repeated sunny summer day in Australia [day–night cycles of incident light (PAR) and temperature]. Monocultures of both marine microalgae were used as reference batch processes. Differences in the autofluorescence of both microalgae enabled the individual measurement, of cell distributions in co-culture, by flow cytometry. The co-cultivation of T. lutea and M. salina in artificial sea water with an inoculation ratio of 1:3 resulted in a balanced biomass production of both microalgae simultaneously with a DHA:EPA ratio of almost 1:1 (26 mg_DHA g_CDW⁻¹, and 23 mg_EPA g_CDW⁻¹, respectively) at harvest after depletion of the initially added fertilizer. Surprisingly, more microalgae biomass was produced within 8 days in co-cultivation with an increase in the cell dry weight (CDW) concentration by 31%, compared to the monocultures with the same amount of light and fertilizer. What is more, DHA-content of the microalgae biomass was enhanced by 33% in the co-culture, whereas EPA-content remained unchanged compared to the monocultures. Graphical Abstract

Studies on microalgal lipid production as a sustainable feedstock for biofuels and chemicals are scarce, particularly those on applying open thin-layer cascade (TLC) photobioreactors under dynamic diurnal conditions. Continuous lipid production with Microchloropsis salina was studied in scalable TLC photobioreactors at 50 m² pilot scale, applying a physically simulated Mediterranean summer climate. A cascade of two serially connected TLC reactors was applied, promoting biomass growth under nutrient-replete conditions in the first reactor, while inducing the accumulation of lipids via nitrogen limitation in the second reactor. Up to 4.1 g L⁻¹ of lipids were continuously produced at productivities of up to 0.27 g L⁻¹ d⁻¹ (1.8 g m² d⁻¹) at a mean hydraulic residence time of 2.5 d in the first reactor and 20 d in the second reactor. Coupling mass balances with the kinetics of microalgal growth and lipid formation enabled the simulation of phototrophic process performances of M. salina in TLC reactors in batch and continuous operation at the climate conditions studied. This study demonstrates the scalability of continuous microalgal lipid production in TLC reactors with M. salina and provides a TLC reactor model for the realistic simulation of microalgae lipid production processes after re-identification of the model parameters if other microalgae and/or varying climate conditions are applied.

Marine microalgae offer a sustainable alternative source for the human diet’s essential omega-3-fatty acids, including docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5). However, none of them can produce DHA and EPA in a nutritionally balanced ratio of 1:1. As shown recently, the phototrophic co-cultivation of the golden-brown microalgae Tisochrysis lutea (DHA producer) with the green microalgae Microchloropsis salina (EPA producer) can provide microalgae biomass with a balanced DHA-to-EPA ratio with increased productivity compared to monocultures. This study evaluates whether other golden-brown (Isochrysis galbana) and green microalgae (Nannochloropsis oceanica, Microchloropsis gaditana) can enable the phototrophic batch production of omega-3 fatty acids in a nutritionally balanced ratio in co-culture. All co-cultivations applying a physically dynamic climate simulation of a repeated sunny summer day in Australia in LED-illuminated flat-plate gas lift photobioreactors resulted in increased biomass concentrations compared to their respective monocultures, achieving balanced DHA-to-EPA ratios of almost 1:1. Using urea instead of nitrate as a nitrogen source increased the EPA content by up to 80% in all co-cultures. Light spectra measurements on the light-adverted side of the photobioreactor showed that increased biomass concentrations in co-cultures could have been related to enhanced light use due to the utilization of different wavelengths of the two microalgae strains, especially with the use of green light (500–580 nm) primarily by golden-brown microalgae (I. galbana) and orange light (600–620 nm) predominantly used by green microalgae (N. oceanica). Phototrophic co-cultivation processes thus promise higher areal biomass yields if microalgae are combined with complimentary light-harvesting features.

Marine microalgae from the genus Nannochloropsis are promising candidates for the photoautotrophic production of eicosapentaenoic acid (EPA, C20:5), a polyunsaturated fatty acid known for its numerous health benefits. A recent study demonstrated that Microchloropsis salina can accumulate high amounts of EPA when cultivated in flat-plate gas-lift photobioreactors. This study aimed to characterize an alternative strain, Nannochloropsis oceanica, and compare its biomass and EPA productivity to M. salina. Applying simulated dynamic climate conditions of a repeated sunny summer day in Eastern Australia, N. oceanica was cultivated in LED-illuminated flat-plate gas-lift photobioreactors. The results showed significantly higher biomass growth and EPA contents compared to M. salina. An EPA productivity of 33.0 ± 0.6 mg_EPA L⁻¹ d⁻¹ has been achieved in batch processes with N. oceanica. Scaling up the photoautotrophic process to 8 m² thin-layer cascade photobioreactors resulted in doubled concentrations of N. oceanica biomass compared to laboratory-scale batch processes. This improvement was likely due to the reduced fluid layer depth, which enhanced light availability to the microalgal cells. Using urea instead of nitrate as a nitrogen source further improved the EPA production of N. oceanica in thin-layer cascade photobioreactors, achieving CDW concentrations of up to 17.7 g L⁻¹ and thus a high EPA concentration of 843 mg L⁻¹. These findings highlight N. oceanica as an alternative to M. salina for sustainable EPA production, offering potential for further industrial applications.

Marine microalgae have received much attention as a sustainable source of the two health beneficial omega-3-fatty acids docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5). However, photoautotrophic monocultures of microalgae can only produce either DHA or EPA enriched biomass. An alternative may be the photoautotrophic co-cultivation of Tisochrysis lutea as DHA-producer with Microchloropsis salina for simultaneous EPA production to obtain EPA- and DHA-rich microalgae biomass in a nutritionally balanced ratio. Photoautotrophic co-cultivation processes of T. lutea and M. salina were studied, applying scalable and fully controlled lab-scale gas-lift flat-plate photobioreactors with LED illumination for dynamic climate simulation of a repeated sunny summer day in Australia (day-night cycles of incident light (PAR) and temperature). Monocultures of both marine microalgae were used as reference batch processes. Differences in the autofluorescence of both microalgae enabled the individual measurement, of cell distributions in co-culture, by flow cytometry. The co-cultivation of T. lutea and M. salina in artificial sea water with an inoculation ratio of 1:3 resulted in a balanced biomass production of both microalgae simultaneously with a DHA:EPA ratio of almost 1:1 (26 mg_DHA g_CDW⁻¹, and 23 mg_EPA g_CDW⁻¹, respectively) at harvest after depletion of the initially added fertilizer. Surprisingly, more microalgae biomass was produced within 8 days in co-cultivation with an increase in the cell dry weight (CDW) concentration by 31%, compared to the monocultures with the same amount of light and fertilizer. What is more, DHA-content of the microalgae biomass was enhanced by 33% in the co-culture, whereas EPA-content remained unchanged compared to the monocultures.