- Measurement of Ascorbic Acid and Glutathione Content in Cyanobacterium Synechocystis sp. PCC 6803.
Measurement of Ascorbic Acid and Glutathione Content in Cyanobacterium Synechocystis sp. PCC 6803.
Bio-protocol (2021-03-05)
Anabella Aguilera, Charlotte Steelheart, Matías Alegre, Federico Berdun, Graciela Salerno, Carlos Bartoli, Gabriela Pagnussat, María Victoria Martin
PMID33659454
摘要
Ascorbic acid (AsA) and gluthathione (GSH) are two key components of the antioxidant machinery of eukaryotic and prokaryotic cells. The cyanobacterium Synechocystis sp. PCC 6803 presents both compounds in different concentrations (AsA, 20-100 μM and GSH, 2-5 mM). Therefore, it is important to have precise and sensitive methods to determine the redox status in the cell and to detect variations in this antioxidants. In this protocol, we describe an improved method to estimate the content of both antioxidants (in their reduced and oxidized forms) from the same sample obtained from liquid cultures of Synechocystis sp. PCC 6803.
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Sigma-Aldrich
5,5'-二硫代双(2-硝基苯甲酸), ≥98%, BioReagent, suitable for determination of sulfhydryl groups
Sigma-Aldrich
谷胱甘肽还原酶 来源于面包酵母(酿酒酵母), ammonium sulfate suspension, 100-300 units/mg protein (biuret)
酸洗玻璃珠, 150-212 μm (70-100 U.S. sieve)