Rational protein design is a great challenge to our understanding of protein structure and physical chemistry. It has been widely used in numerous fields, including synthetic biology. Rational design is the most significant approach of protein engineering based on site-directed mutagenesis. The strategy is based on the relating structure to function, frequently via molecular modeling techniques. Rational design requires the knowledge of the amino acid sequence, 3D structure and function of the target protein. This method provides controllable amino acid sequence changes such as insertion, deletion, substitution, exchange of secondary structural elements, exchange of the entire domain, or fusion of enzymes to create new proteins.
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