Microalgae adaptation as a strategy to recycle the aqueous phase from hydrothermal liquefaction

Hydrothermal liquefaction (HTL) produces bio-crude oil from wet algae along with an aqueous phase (AP). This effluent contains minerals that can be reused for cultivating new microalgae but whose utility remains limited due to the presence of inhibitors. Reduced photosynthetic performance, growth, and null lipid accumulation were observed in wild-type Chlorella vulgaris NIES 227 cultivated in AP (1/200). Adaptive laboratory evolution was studied by batch transfers and turbidostat mode. Both methods effectively counterbalanced AP toxicity and restored the fitness of the microalgae. After adaptation, a higher AP addition was achieved, from 1/600 to 1/200, without inhibition. As compared with the wild typein control medium (0.261 g/L/d), both adapted-strains maintained competitive productivity (0.310 and 0.258 g/L/d) of lipid-rich biomass (37 %-56 %). The improved tolerance of the adapted strains persisted after the removal of AP and under axenic conditions. Adaptive laboratory evolution is suggested for AP reutilization in the algae production process.