Abstract
Directed evolution is a powerful tool for engineering protein function. The process of directed evolution involves iterative rounds of sequence diversification followed by assaying activity of variants and selection. The range of sequence variants and linked activities generated in the course of an evolution are a rich information source for investigating relationships between sequence and function. Key residue positions determining protein function, combinatorial contributors to activity and even potential functional mechanisms have been revealed in directed evolutions. The recent application of high throughput sequencing substantially increases the information that can be retrieved from directed evolution experiments. Combined with computational analysis this additional sequence information has allowed high-resolution analysis of individual residue contributions to activity. These developments promise to significantly enhance the depth of insight that experimental evolution provides into mechanisms of protein function.
Highlights
Directed evolution is one of the most effective strategies currently available for modifying functional activities of proteins.[1,2,3] most directed evolution studies have been aimed at creating or modifying protein function, this approach can provide significant insights into fundamental aspects of protein binding, enzyme catalysis and structure–function relationships
Key residue positions determining protein function, combinatorial contributors to activity and even potential functional mechanisms have been revealed in directed evolutions
The recent application of high throughput sequencing substantially increases the information that can be retrieved from directed evolution experiments
Summary
Directed evolution is one of the most effective strategies currently available for modifying functional activities of proteins.[1,2,3] most directed evolution studies have been aimed at creating or modifying protein function, this approach can provide significant insights into fundamental aspects of protein binding, enzyme catalysis and structure–function relationships. The ability to sequence large numbers of variants, both intermediates generated during the evolution and final evolved variants, together with computational analysis tools will help to better identify combinatorial contributors to function and provide more information about how these residues work together.
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