*3.2. Culture Conditions*

### 3.2.1. Autotrophic Culture of *T. suecica*

The microalgae *T. suecica* was cultured in 100-mL glass photobioreactors with a diameter of 3.0 cm, with constant temperature (21 ◦C). The photoperiod was adjusted to 12 h of light (irradiance 165 μmol photons m<sup>−</sup><sup>2</sup> s<sup>−</sup>1) and 12h of darkness (12:12) and was maintained in agitation with a constant air bubbling system in order to avoid microalgae sedimentation and achieving a homogeneous

distribution of nutrients and irradiance in each cell. The culture was inoculated with a concentration of 6 × 10<sup>5</sup> cells mL−<sup>1</sup> and adjusted to 35 ‰ salinity. The culture system was through a batch culture, with the addition of the culture medium F/2 [81]. The culture was volumetrically scaled to 10 L and was kept in a stationary phase. All the experiments were carried out in triplicate.

### 3.2.2. Heterotrophic Culture of *T. suecica*

The adaptation of an autotrophic to a heterotrophic culture of *T. suecica* was carried out in 100-mL glass photobioreactors with a diameter of 3.0 cm and constant temperature (21 ◦C). A batch culture system was started. The reduction of the photoperiod illumination time was progressive (light:dark) 12:12 (288 h), 8:16 (228 h), 4:20 (240 h), 3:21 (156 h), 2:22 (168 h) and 0:24. The culture medium used was F/2 supplemented with penicillin/streptomycin 0.1% *<sup>v</sup>*/*<sup>v</sup>*, with an optimal glucose dosage of5gL−<sup>1</sup> in darkness condition. The culture was maintained with constant agitation by means of an air bubbling system, avoiding the sedimentation of the microalgae and achieving a homogeneous distribution of nutrients. Once the cultures were adapted to heterotrophy, they were volumetrically scaled to reach 2 L. All the experiments were carried out in triplicate.
