Extraction of Recombinant Protein From Bacteria
The use of bacteria for overexpression of recombinant proteins is still a popular choice because of lower cost and higher yields when compared to other expression systems (1,2), but problems can arise in the recovery of soluble functionally active protein. In some cases, secretion of recombinant proteins by bacteria into the media has eliminated the need to lyse the cells, but most situations still require lysis of the bacterial cell wall in order to extract the recombinant protein product. A number of methods based on enzymatic methods and mechanical means are available for breaking open the bacterial cell wall, and the choice will depend on the scale of the process (3,4). Enzymatic methods include lysozyme hydrolysis, which cleaves the glucosidic linkages in the bacterial cell-wall polysaccharide. The inner cytoplasmic membrane can then be disrupted easily by detergents, osmotic pressure, or mechanical methods.
KeywordsSodium Dodecyl Sulfate Recombinant Protein Inclusion Body Bacterial Cell Wall Recombinant Protein Product
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