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Perpetua: The Journal of Undergraduate Research at UAH

College

College of Science

Department

Biological Sciences

Abstract

Protein purification requires an intensive multi-step process that is customized to every protein’s characteristic for optimal results. In this study, we looked at the purification process of Inorganic Pyrophosphatase (IPPase), an essential protein found in all living organisms. The commonality of the protein does not veil the fact that the mechanism by which IPPase is hydrolyzed into two orthophosphates is not fully understood. To comprehend the mechanism, recombinant mutants of the conservative region in IPPase were expressed. However, to observe a possible change in structure of the recombinant mutants with crystallization, pure concentrated protein is needed. Therefore, protein purification is crucial. During this purification process, three characteristics of IPPase are manipulated to create an acceptable purification: IPPase’s thermophilic nature, its isoelectric point, and the 21 kDa size of the monomer. With the perfection of functioning protocol, steps can begin to be taken in the studying of the activity of each integral amino acid.

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