Comparison of monocultures and a mixed culture of three Chlorellaceae strains to optimize biomass production and biochemical content in microalgae grown in a greenhouse

Light and temperature are important environmental conditions affecting microalgal growth in outdoor culture. It is essential to evaluate microalgae strains growing under outdoor conditions where they are subjected to variable environmental parameters. The present study investigated three Chlorellaceae strains (Micractinium sp. MACC-728, Chlorella sorokiniana MACC-438, and C. sorokiniana MACC-452) and a mixed culture combining these three strains. Cultures were grown in 2-L bioreactors in a greenhouse over 3 months to assess the effects of high temperature and light on their growth, macromolecule content, and antioxidant and plant-stimulating bioactivities. The most influential environmental parameters on growth were average air temperature and the sum of photosynthetically active radiation, followed by maximum air temperature. The most affected growth parameter was daily change in cell number. Chlorella sorokiniana MACC-438 produced the lowest biomass and was most affected by the high temperature and light conditions. Micractinium sp. produced the highest biomass and was least affected, suggesting it was the most suitable strain for outdoor cultivation. The mixed Chlorellaceae culture performed well in biomass production, exceeding C. sorokiniana monocultures but significantly underyielding in lipid content. Antioxidant activity and the root-stimulating activity varied with strain and culture age. Micractinium sp. had the highest but most variable antioxidant and plant-stimulating activity. Bioactivity in the mixed culture was more consistent, remaining high regardless of culture age and environmental conditions. Thus, mixed cultures of productive strains could be a useful strategy to ensure stable and high-quality biomass production in outdoor cultivation with fluctuating environmental conditions.