Carbon distribution and metabolism mechanism of a novel mixotrophic Chlorella in municipal wastewater

Conventional wastewater treatment technologies primarily convert complex organic matter into dissolved inorganic carbon (DIC) and a more difficult gaseous state CO₂. Most microalgae species can photosynthetically assimilate above inorganic carbon, but their heterotrophic metabolic processes often dominate in glucose-mediated mixotrophy. Herein, we investigated the carbon-fixing metabolic pathways of Chlorella sp. MIHQ61 in municipal wastewater containing complex carbon sources. The total carbon removal (73.0 %) peaked on the 6th day, and DIC removal exceeded 50.0 % as the carbon migrating amount from municipal wastewater into the microalgal cells peaked. The glucose and NaHCO₃ combination promoted both autotrophic and heterotrophic metabolism. Headspace CO₂ emission, enzyme activity and central carbon metabolism results implied heterotrophic metabolism occurred more actively in the early stage and autotrophic metabolism dominated late stage. Redefined mixotrophic carbon allocation by revealing time-dependent autotrophic/heterotrophic interplay. Carbon distribution and mixotrophic mechanism provided new thinking on how to utilize microalgae and wastewater resource.