## 2 Materials and Methods
### Biochemical Composition  
Total lipid content was determined according to the Bligh and Dyer [25] method with some modifications [26]. Briefly, approximately 20 mg of freeze-dried biomass was extracted with a mixture of chloroform, methanol, and water (1:2:0.8, _v:v:v_) and homogenized for 3 minutes in a mixer mill (MM400, Retsch GmbH, Retsch-Alle 1-5, 42781 Haan, Germany) with glass beads at 30 Hz. The chloroform layer, separated by centrifugation, was transferred to new vessels. Then, 0.7 mL were pipetted to preweighed tubes and evaporated at 60 \({}^{\circ}\)C. The resulting dried residue was weighed (Sartorius Lab Instruments GmbH & Co. 370/70 Gottingen, Germany-MSA36S-000-DH) and compared with the initial weight to provide an accurate result of the lipid fraction. Protein content was calculated by multiplying the N content with the standard conversion factor of 6.25 [27] after CHN analysis using a Vario EL III (Vario EL, Elementar Analyser System, GmbH, Hanau, Germany). The ash content was determined by the biomass's weight difference before and after combustion. Biomass was heated at 550 \({}^{\circ}\)C for 8 h using a furnace (LE062K17N1, Nabertherm, Lilienthal, Germany). Carbohydrates were determined by differences from other macronutrients. Fatty acid methyl esters (FAME) were determined according to [PERSON] and [PERSON] [28] protocol, modified by [PERSON] et al. [26]. Briefly, 20 mg of freeze-dried biomass was added to 1.5 mL of methanol/acetyl chloride (20:1 _v:v_) and homogenized for 3 minutes in a mixer mill (MM400, Retsch GmbH, Retsch-Alle 1-5, 42781 Haan, Germany) with glass beads, at 30 Hz. Then, 1 mL of \(n\)-hexane was added to the derivatization vessels and placed in a water bath (70 \({}^{\circ}\)C for 60 min). Thereafter, water and \(n\)-hexane (1:4, _v:v_) were added and homogenized in a vortex. The hexane fraction, obtained by centrifugation, was transferred to new tubes, and anhydrous sodium sulfate (Na\({}_{2}\)SO\({}_{4}\)) was added to the extracts. After filtration, samples were evaporated under nitrogen gas flow. The pellet was resuspended in 0.5 mL of gas chromatography-grade hexane and analyzed in a GC-MS analyzer (Bruker SCION 456/GC, SCION TQ MS) equipped with a ZB-5 MS capillary column (30 m \(\times\) 0.25 mm of internal diameter, 0.25 \(\upmu\)m of film thickness, Phenomenex), using helium as the carrier gas. The temperature was set to 60 \({}^{\circ}\)C for 1 min, then 30 \({}^{\circ}\)C min\({}^{-1}\) to 120 \({}^{\circ}\)C, 5 \({}^{\circ}\)C min\({}^{-1}\) to 250 \({}^{\circ}\)C and finally increased to 20 \({}^{\circ}\)C min\({}^{-1}\) up to 300 \({}^{\circ}\)C, with the injection temperature being 300 \({}^{\circ}\)C as well. To establish the calibration curves, five different concentrations of the standards Supelco(r) 37 component FAME Mix 41 (Sigma-Aldrich, Sintra, Portugal) were analyzed for further identification and quantification of FAME. For chlorophylls \(a\) and \(b\) quantification, 10 mg of freeze-dried biomass were weighed, 6 mL acetone was added and homogenized for 10 min with glass beads in a vortex. The samples were centrifuged (HERMLE Z300, New York, NY, USA) at 2500\(\times\)\(g\) for 10 min until complete loss of the pellet or the supernatant color. The supernatant was analyzed using a spectrophotometer (Genesys 10S UV-VIS, Waltham, MA, USA) in the scanning spectrum (380 to 700 nm), and the resultant data by the Excel Add-In Solver on Windows 2013. The content of carotenoids was determined based on [PERSON] et al. [29]. Briefly, 10 mg of freeze-dried biomass was resuspended in methanol containing 0.03% of butylhydroxy-toluene and homogenized for 3 minutes in a mixer mill (MM400, Retsch GmbH, Retsch-Alle 1-5, 42781 Haan, Germany), with glass beads and one tungsten bead, at 30 Hz, until complete loss of the pellet or the supernatant color. Then, the centrifuged extract was completely dried under nitrogen flow, and the pellet was resuspended in high-performance liquid chromatography (HPLC)-grade methanol and filtered (0.22 \(\upmu\)m) into amber vials.