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.