Thermosynechococcus sp. CL-1 (TCL-1) has a high potency to utilize CO2 under extreme conditions including high temperature, alkaline condition, and the occurrence of 17β-estradiol (E2). In this study, TCL-1 cultivation with E2 addition in the range of 0–20 mg/L was combined with various growth arrangements (light intensity and dissolved inorganic nitrogen/DIN level). After 120 h cultivation, the 1.0 mg/L E2, 200 µmol photons/m2/s light intensity, and 5.8 mM available nitrogen performed the best growth with 4.58 ± 0.18 mg/L/h biomass productivity, 94.9 ± 3.3% total estrogen removal, and 11.41 ± 0.11 mg/L/h CO2 fixation rate. Estrogen degradation was mainly carried out by biodegradation route which started from E2 conversion into estrone/E1 and with only 4–6% influence from the abiotic factors. Compared with the accumulated zeaxanthin, β-carotene was dominantly generated with a productivity of 0.043 ± 0.019 mg/L/h. Therefore, TCL-1 cultivation is an efficient strategy for simultaneous CO2 fixation, estrogen removal, and carotenoid accumulation as valuable byproducts.
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