Electrophoretic and Chromatographic Analysis of Photosynthetic Pigments from Blue-Green Algae

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Electrophoretic and Chromatographic Analysis of Photosynthetic Pigments from Blue-Green Algae

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Cyanobactera, also known as blue-green algae, obtain their energy by photosynthesis using sunlight as their energy source. These organisms have been considered to be the oldest and the most important bacteria on the earth. It is believed that they were responsible for the initial oxygenation of the earth’s atmosphere through photosynthesis and it is also felt they were the precursors to the chloroplasts that are found in true algae and plants. There are two classes of photosynthetic pigments in Cyanobactera. The first class contains water-soluble proteins and the major protein is called Phycocyanin, which is blue. The other classes of photosynthetic pigments that include the carotinoids and chlorophylls are small molecular weight molecules and are insoluble in water but soluble in organic solvents such as alcohol. In this laboratory exercise, 8 groups of students isolate and characterize both groups of pigments. In part A of this exercise, students prepare a water-soluble extract from blue green algae and show that it contains the single major protein Phycocyanin by electrophoresis as shown in the gel below. They also determine the charge of this protein by comparing its electrophoretic mobility to the mobilities of dyes with known charges. In part B, they prepare an alcohol extract and analyze the smaller alcohol soluble pigments by thin layer chromatography in order to identify the chlorophylls and major carotenoid pigments. The results of this two-part study give students practical hands-on experience with isolation of components from cells as well as electrophoresis and thin layer chromatography and introduces them to one of the most important organisms on the earth.

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