![]() ![]() It is a well-known fact that the full-length proteins are difficult to be expressed in E. The chaotropic agents increase the entropy by interfering with the noncovalent forces like hydrogen bonding, Vander wall forces, and hydrophobic effects. This is also achieved by using the mild conditions to recover the functional active form (soluble) by using different concentrations of Chaotropic agents like urea and guanidine hydrochloride (GdnHcL) which significantly improve the yield of a recombinant protein. The extracted proteins from the inclusion bodies are used to study the conformation changes by using denaturants. coli) are partially folded intermediates which form the aggregates (insoluble form) known as inclusion bodies. In many cases, recombinant proteins produced in Escherichia coli (E. After isolation, proteins are insoluble in their innate state and must be denatured to solubilization. The polydispersity of the protein sample or binding of ligand or size of proteins can be characterized by using the Dynamic Light Scattering (DLS). Proteins are often associated with other proteins which lead them to form an oligomeric complex. Proteins are large bio-molecules present in their native state which perform the various biological functions. ![]() The aim of this review is to have a profound discussion on these key factors and analyze them in relation to both aspects purification and crystallization and provide a fruitful advice for boosting the production rate of protein and crystallization effectively. In this review, the key factors such as the expression system, Affinity –Tags, solubility, reducing or oxidizing environment, denaturing agents, concentration of precipitant, concentration of protein, ionic strength, isoelectric point and pH, temperature, additives, ligands, presence of substrates, coenzymes, mutation, that affect both protein purification and crystallization are discussed in detail. However, the purity and rate of crystallization success still needs to be improved. The protein solubility is achieved by the Site-directed mutagenesis that generates the hydrophobic to hydrophilic mutations. The strategies developed for protein solubility and crystallization have improved the protein production and provide high-resolution crystals for structural studies. Protein purification and crystallization problems have been noticed for many years. ![]()
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