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Crude proteins and pigments were extracted from different microalgae strains, both marine and freshwater. The effectiveness of enzymatic pre-treatment prior to protein extraction was evaluated and compared to conventional techniques, including ultrasonication and high-pressure water extraction. Enzymatic pre-treatment was chosen as it could be carried out at mild shear conditions and does not subject the proteins to high temperatures, as with the ultrasonication approach. Using enzymatic pre-treatment, the extracted proteins yields of all tested microalgae strains were approximately 0.7 mg per mg of dry cell weight. These values were comparable to those achieved using a commercial lytic kit. Ultrasonication was not very effective for proteins extraction from Chlorella sp., and the extracted proteins yields did not exceed 0.4 mg per mg of dry cell weight. For other strains, similar yields were achieved by both treatment methods. The time-course effect of enzymatic incubation on the proteins extraction efficiency was more evident using laccase compared to lysozyme, which suggested that the former enzyme has a slower rate of cell disruption. The crude extracted proteins were fractionated using an ion exchange resin and were analyzed by the electrophoresis technique. They were further tested for their antioxidant activity, the highest of which was about 60% from Nannochloropsis sp. The total phenolic contents in the selected strains were also determined, with Chlorella sp. showing the highest content reaching 17 mg/g. Lysozyme was also found to enhance the extraction of pigments, with Chlorella sp. showing the highest pigments contents of 16.02, 4.59 and 5.22 mg/g of chlorophyll a, chlorophyll b and total carotenoids, respectively.

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The present study investigated the growth, harvesting, biocrude conversion, and recycling of the HTL aqueous phase for one self-settling (i.e., Chlorocystis sp.) and another non-settling (i.e., Picochlorum sp.) marine microalgae. Both the strains were grown simultaneously in 2 identical 25,000-L raceway ponds in the Qatari desert. The cell size of Picochlorum sp. was small (2-3 µm), and its biomass was harvested using a centrifuge. Cells of Chlorocystis sp. (6-9 µm) formed flocs that settled spontaneously in a sedimentation chamber. Harvested biomass of these two strains was then converted to biocrude oil, using a 500-mL Parr reactor. The biocrude yield of Picochlorum sp. and Chlorocystis sp. was 39.6 ± 1.15% and 34.8 ± 1.65%, respectively. The energy content of the biocrude oil was 32.78 and 33.38 MJ/kg for Chlorocystis sp. and Picochlorum sp., respectively. Both the strains were capable of efficiently utilizing more than 95% nitrogen of the HTL aqueous phase. Although lower biocrude yield was obtained from Chlorocystis sp., compared to Picochlorum sp., harvesting of Chlorocystis sp. would require much lower energy compared to Picochlorum sp. Therefore, a self-settling microalgae (e.g., Chlorocystis sp.) could potentially be a better candidate, over non-settling microalgae, for producing biofuel feedstock.