The green- or red-stage D3–1 cells (50 mL culture) grown under SCC for 1 or 3 months were harvested and freeze-dried using a freeze dryer (FZ-Compact: Labconco Co. Ltd., Kansas City, USA). The DCW was measured, and the samples were subjected to cell disruption with glass beads (φ = 1 ± 0.1 mm) and a mixture of diethyl ether:chloroform:methanol (400 μL, 1:2:1) using Vortex-Genie 2 (Scientific Industries, Inc., New York, USA). The supernatants were obtained via centrifugation (8,000 × g, 10 min). This extraction was repeated four times and approximately 1.6 mL of the extract was prepared. Notably, the cell disruption rate was measured using an optical microscope. These rates were considered to calculate the yield of the extracts. The resultant samples were subjected to a rotary evaporator (V-850; BUCHI, Flawil, Switzerland), and the dried extracts were weighed (approximately 10−30 mg per sample) and stored at −30 °C until use.
Fz compact
Manufactured by Labconco
Sourced in United States
The FZ-Compact is a compact freeze dryer designed for laboratory applications. It offers a freeze-drying solution for small sample volumes. The core function of the FZ-Compact is to remove water from samples through the process of sublimation, preserving the sample's structure and composition.
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Lab products found in correlation
4 protocols using fz compact
1
Extraction and Quantification of Microalgal Lipids
2
Purification and Characterization of Phycocyanin
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The aqueous supernatant containing phycocyanin described in the former section was mixed with 0.5% (w/v) activated carbon and stirred at room temperature for 15 min. Subsequently, the purified phycocyanin aqueous fraction was obtained via centrifugation at 4300×g for 30 min (10 °C). The phycocyanin purity was determined using the following formula [30 (link)] in which OD280 indicates the absorbance of total proteins and OD618 indicates the absorbance of phycocyanin.
The aqueous solution of phycocyanin in a 50 mL centrifuge tube was freeze-dried using a vacuum freeze-dryer (FZ-Compact; Labconco Co. Ltd.), and the resulting phycocyanin powder was weighed directly. The purity and molecular weight of phycocyanin were further assessed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) [31 (link)] as follows. SDS-PAGE loading buffer and the aqueous solution of phycocyanin (4 mg mL− 1) were mixed and heated at 95 °C for 2 min, following which 25 μL of the resultants were subjected to electrophoresis on a 12.5% SDS-polyacrylamide gel. After electrophoresis, the gel was stained with Coomassie Brilliant Blue R250 (Nacalai Tesque, Tokyo, Japan).
The aqueous solution of phycocyanin in a 50 mL centrifuge tube was freeze-dried using a vacuum freeze-dryer (FZ-Compact; Labconco Co. Ltd.), and the resulting phycocyanin powder was weighed directly. The purity and molecular weight of phycocyanin were further assessed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) [31 (link)] as follows. SDS-PAGE loading buffer and the aqueous solution of phycocyanin (4 mg mL− 1) were mixed and heated at 95 °C for 2 min, following which 25 μL of the resultants were subjected to electrophoresis on a 12.5% SDS-polyacrylamide gel. After electrophoresis, the gel was stained with Coomassie Brilliant Blue R250 (Nacalai Tesque, Tokyo, Japan).
3
Cyanobacterial Cell Cultivation and Analysis
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A 200 μL aliquot of the cell culture was collected and placed in a well of a 96-well microtiter plate. The absorption spectrum was measured from 250 to 830 nm using a Multiskan Go spectrometer (Thermo Fisher Co. Ltd., Tokyo, Japan). The cyanobacterial cells cultured in the 6 L tank were harvested via centrifugation and freeze-dried using a vacuum freeze dryer (FZ-Compact; Labconco Co. Ltd., Kansas City, USA). The dry cell weight was measured by using electric balance (AY303; Sartorius Co.Ltd., Göttingen, Germany).
4
Measurement of pH, CO2, and Biomass
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The culture pH, dissolved carbon dioxide concentration (DCDC), and biomass (Supplementary Table S1 , Figures 2 , 3 ) were measured. To measure the pH, 50 mL of the supernatant of the BG11 medium or BX1.5 culture was harvested, and the pH was measured (LAQUA F-71: Horiba Co., Ltd., Kyoto, Japan). To measure DCDC, 50 mL of the BG11 medium or BX1.5 culture supernatant was harvested and measured using a portable carbon dioxide gas concentration metre (Model CGP-31, DKK-TOA Co., Ltd., Tokyo, Japan). The 50 mL of BX1.5 culture were harvested, centrifuged (8,000 × g, 10 min, 10°C), and the cell biomass containing the BX1.5 extracellular polysaccharide (bxEPS) was stored at −80°C until use. The cell matter was freeze-dried as DCM (dried cell matter with bxEPS) in a vacuum freeze dryer (FZ-Compact; Labconco Co., Ltd., Kansas City, United States of America) for 72 h. The weight of DCM (DMW) was measured using an electron precision balance (MS603S; METTLER TOLEDO, Tokyo, Japan). Of note, since the supernatant and cells cannot be separated by centrifugation due to the abundant bxEPS production, 20 mL of the cell culture was directly transferred to a 50 mL tube, frozen at −80°C, and lyophilised as the DCM sample.
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