The purpose of this work is to define optimal growth conditions to maximise biomass for batch culture of the cyanobacterium Arthrospira maxima and the microalgae Chlorella vulgaris, Isochrysis galbana and Nannochloropsis gaditana. Thus, we study the effect of three variables on cell growth: i.e., inoculum:culture medium volume ratio (5:45, 10:40, 15:35 and 20:30 mL:mL), light:dark photoperiod (8:16, 12:12 and 16:8 h) and type of culture medium, including both synthetic media (Guillard’s F/2 and Walne’s) and wastewaters. The results showed that the initial inoculum:culture medium volume ratio, within the range 5:45 to 20:30, did not affect the amount of biomass at the end of the growth (14 days), whereas high (18 h) or low (6 h) number of hours of daily light was important for cell growth. The contribution of nutrients from different culture media could increase the growth rate of the different species. A. maxima was favoured in seawater enriched with Guillard’s F/2 as well as C. vulgaris and N. gaditana, but in freshwater medium. I. galbana had the greatest growth in the marine environment enriched with Walne’s media. Nitrogen was the limiting nutrient for growth at the end of the exponential phase of growth for C. vulgaris and N. gaditana, while iron was for A. maxima and I. galbana. The growth in different synthetic culture media also determines the biochemical composition of each of the microalgae. All species demonstrated their capability to grow in effluents from a wastewater treatment plant and they efficiently consume nitrogen, especially the three microalga species.

The production of fatty acids ethyl esters (FAEEs) to be used as biodiesel from oleaginous microalgae shows great opportunities as an attractive source for the production of renewable fuels without competing with human food. To ensure the economic viability and environmental sustainability of the microbial biomass as a raw material, the integration of its production and transformation into the biorefinery concept is required. In the present work, lipids from wet Isochrysis galbana microalga were extracted with ethyl acetate with and without drying the microalgal biomass (dry and wet extraction method, respectively). Then, FAEEs were produced by lipase-catalyzed transesterification and esterification of the extracted lipids with ethanol using lipase B from Candida antarctica (CALB) and Pseudomonas cepacia (PC) lipase supported on SBA-15 mesoporous silica functionalized with amino groups. The conversion to FAEEs with CALB (97 and 85.5 mol% for dry and wet extraction, respectively) and PC (91 and 87 mol%) biocatalysts reached higher values than those obtained with commercial Novozym 435 (75 and 69.5 mol%). Due to the heterogeneous nature of the composition of microalgae lipids, mixtures with different CALB:PC biocatalyst ratio were used to improve conversion of wet-extracted lipids. The results showed that a 25:75 combi-lipase produced a significantly higher conversion to FAEEs (97.2 mol%) than those produced by each biocatalyst independently from wet-extracted lipids and similar ones than those obtained by each lipase from the dry extraction method. Therefore, that optimized combi-lipase biocatalyst, along with achieving the highest conversion to FAEEs, would allow improving viability of a biorefinery since biodiesel production could be performed without the energy-intensive step of biomass drying.

The yield and composition of the biocrude obtained by hydrothermal liquefaction (HTL) of Nannocloropsis gaditana using heterogeneous catalysts were evaluated. The catalysts were based on metal oxides (CaO, CeO₂, La₂O₃, MnO₂, and Al₂O₃). The reactions were performed in a batch autoclave reactor at 320 °C for 10 min with a 1:10 (wt/wt) microalga:water ratio. These catalysts increased the yield of the liquefaction phase (from 94.14 ± 0.30 wt% for La₂O₃ to 99.49 ± 0.11 wt% for MnO₂) as compared with the thermal reaction (92.60 ± 1.20 wt%). Consequently, the biocrude yields also raised in the metal oxides catalysed HTL, showing values remarkably higher for the CaO (49.73 ± 0.9 wt%) in comparison to the HTL without catalyst (42.60 ± 0.70 wt%). The N and O content of the biocrude obtained from non-catalytic HTL were 6.11 ± 0.02 wt% and 10.50 ± 0.50 wt%, respectively. In this sense, the use of the metal oxides decreased the N content of the biocrude (4.62 ± 0.15–5.45 ± 0.11 wt%), although, they kept constant or increased its O content (11.39 ± 2.06–21.68 ± 0.03 wt%). This study shows that CaO, CeO₂ and Al₂O₃ can be promising catalysts based on the remarkable amount of biocrude, the highest values of C, H, heating value, energy recovery, and the lowest content of N, O and S.

Phaeodactylum tricornutum is an interesting source of biomass to produce biocrude by hydrothermal liquefaction (HTL). Its biochemical composition, along with its biomass productivity, can be modulated according to this specific application by varying the photoperiod, the addition of CO₂ or the variation of the initial nitrate concentration. The lab-scale culture allowed the production of a P. tricornutum biomass with high biomass and lipid productivities using a 18:6 h light:dark photoperiod and a specific CO₂ injection. An initial concentration of nitrates (11.8 mM) in the culture was also essential for the growth of this species at the lab scale. The biomass generated in the scale-up photoreactor had acceptable biomass and lipid productivities, although the values were higher in the biomass cultivated at the lab scale because of the difficulty for the light to reach all cells, making the cells unable to develop and hindering their growth. The biocrudes from a 90-L cultivated microalga (B-90L) showed lower yields than the ones obtained from the biomass cultivated at the lab scale (B-1L) because of the lower lipid and high ash contents in this biomass. However, the culture scaling-up did not affect significantly the heteroatom concentrations in the biocrudes. A larger-scale culture is recommended to produce a biocrude to be used as biofuel after a post-hydrotreatment stage.

The production of fatty acids ethyl esters (FAEEs) to be used as biodiesel from oleaginous microalgae shows great opportunities as an attractive source for the production of renewable fuels without competing with human food. To ensure the economic viability and environmental sustainability of the microbial biomass as a raw material, the integration of its production and transformation into the biorefinery concept is required. In the present work, lipids from wet Isochrysis galbana microalga were extracted with ethyl acetate with and without drying the microalgal biomass (dry and wet extraction method, respectively). Then, FAEEs were produced by lipase-catalyzed transesterification and esterification of the extracted lipids with ethanol using lipase B from Candida antarctica (CALB) and Pseudomonas cepacia (PC) lipase supported on SBA-15 mesoporous silica functionalized with amino groups. The conversion to FAEEs with CALB (97 and 85.5 mol% for dry and wet extraction, respectively) and PS (91 and 87 mol%) biocatalysts reached higher values than those obtained with commercial Novozym 435 (75 and 69.5 mol%). Due to the heterogeneous nature of the composition of microalgae lipids, mixtures with different CALB:PC biocatalyst ratio were used to improve conversion of wet-extracted lipids. The results showed that a 25:75 combi-lipase produced a significantly higher conversion to FAEEs (97.2 mol%) than those produced by each biocatalyst independently from wet-extracted lipids and similar ones than those obtained by each lipase from the dry extraction method. Therefore, that optimised combi-lipase biocatalyst, along with achieving the highest conversion to FAEEs, would allow improving viability of a biorefinery since biodiesel production could be performed without the energy-intensive step of biomass drying.

The aim of the present work was focused on optimising the yield and quality of the biocrude obtained by hydrothermal liquefaction (HTL) of Nannochloropsis gaditana. Temperature, reaction time and microalga concentration were the variables used to carry out an experimental factorial design with a central composite design. The responses chosen were the biocrude yield and the nitrogen and oxygen content in the biocrude phase. A second-order model was obtained to predict the responses as a function of these variables. Temperature is the most determining factor with a positive influence on biocrude yield. The maximum biocrude yield (42.3 ± 0.8 wt%) was obtained at 320 °C, 10 min of reaction and 10 wt% microalgae concentration, and the nitrogen and oxygen content significantly decreased with respect to their corresponding levels in the initial microalgal biomass. The HHV value of the biocrude was 35.7 MJ/kg. The biocrude was composed of 30% of linear and branched hydrocarbons.