2.2 Extraction of Glutathione, Phycocyanin and SOD from
Microalgae Cells
As shown in Figure 1, Microcystis aeruginosa FACHB-905 was cultured on
days 7, 14, 21, 28, 35, 50, 65, 80 and 95, respectively. The algal
sludge was obtained by centrifugation at 4 ℃ and 8000 rpm for 10 min.
(1).Extraction of glutathione from algae cells: The broken algal cells
were added with 5% sulfosalicylic acid to deproteinize the algal cells
in suspension state. The samples were centrifuged for 15 min and soaked
with 10 mL PBS for 2 h. After centrifugation for 20 min, the supernatant
was GSH extract.
(2). Extraction of phycocyanin from algae cells: Phosphate buffer at 4 ℃
was added to algal mud samples in which Microcystis aeruginosa cells
were infiltrated before 4 ℃. Freeze and thaw three more times. The
disrupted algal cells were suspended in acetone containing 40 mL of TCA
and DTT and precipitated at -20°C for 2 hours. The samples were
centrifuged to discard the supernatant, precipitated and added to the
acetone solution, and repeated 2-3 times to obtain the phycocyanin
extract.
(3). Extraction of SOD from algae cells: The broken algal cells were
crushed in PBS phosphate buffer to infiltrate the algal cells, and the
supernatant was centrifuged to obtain the enzyme extract. The above
samples were extracted with 5 mL of chloroform-ethanol mixture to remove
various proteins. Finally, the precipitate was centrifuged and discarded
to obtain SOD enzyme liquid.