H. lacustris NIES-4141 was provided by the Microbial Culture Collection at the NIES. The autotrophic medium AF-6 was used for culture (working volume, 20 ml). Cells were grown in culture flasks under 14-hour light/10-hour dark cycle illumination (120 to 140 μmol m−2 s−1) at 25°C. After a 5-day preculture in normal AF-6 medium, the cells were transferred to nitrogen-deficient AF-6 medium (without NH4NO3 and NaNO3) at a cell density of 1 × 105 cells/ml to induce the production of astaxanthin. The series of measurements were performed 0 to 5 days after the replacement of the culture medium.

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