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Author Topic: PROTEIN BIOTECHNIQUES- Extractions of Protein  (Read 3702 times)
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Francis Umeoguaju
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« on: April 10, 2010, 08:30:43 am »


Before going into the extraction of protein, one must be sure that the enzyme or protein of interest is present in appreciable quantity in the sample that one is using for the extraction, it will not be reasonable or economically feasible, for instance, to try to extract alpha amylase from beans when it could be obtained in a much larger quantity in germinating Barley. (except beans alpha amylase is specifically required even at that a condition that would enhance the biosynthesis of the enzyme needs to be created in the plant or animal prior to isolation/extraction.)

Another factor to consider is the choice of extraction solvent. Since protein have different solubility in various media, depending on their pI, the idea will be to use a media in which most of the protein of interest will dissolve while excluding much of the contaminating protein and at the same time preserving the native conformation of the protein of interest.

There are various solvent that can be used to extract proteins this includes water, saline, buffer solutions, TCA, etc. 

The choice of a solvent may also be influenced by the physiological state of the protein. Some proteins are easily denatured once they are away from their physiological environment, for such protein, a buffered solution would be necessary for extraction.

Generally to extract protein from plant seeds, the seeds are crushed with cold mortal and pestle and are then blended with suitable extraction solvent. (a blender may also be used for this purpose)  The extracts are filtered through a muslin cloth. The filtrates are then centrifuged to obtain the soluble portion of the extract. The pellet are discarded and the supernatant  which comprise thousands of soluble proteins are then frozen until required.
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