Abstract
AbstractEvolution is essential to the generation of complexity and ultimately life. It relies on the propagation of the properties, traits, and characteristics that allow an organism to survive in a challenging environment. It is evolution that shaped our world over about four billion years by slow and iterative adaptation. While natural evolution based on selection is slow and gradual, directed evolution allows the fast and streamlined optimization of a phenotype under selective conditions. The potential of directed evolution for the discovery and optimization of enzymes is mostly limited by the throughput of the tools and methods available for screening. Over the past twenty years, versatile tools based on droplet microfluidics have been developed to address the need for higher throughput. In this Review, we provide a chronological overview of the intertwined development of microfluidics droplet‐based compartmentalization methods and in vivo directed evolution of enzymes.
Highlights
For enantiospecific ester hydrolysis designed from short peptides,[14] and of a metalloenzyme for olefin metathesis using an expanded nitrobindin variant.[15]
One of the main requirements in directed evolution is linking the activity of a target enzyme to its genetic information, which is essential for screening, selection and evolution campaigns.[21,22]
In the last twenty years, engineering and biochemistry research groups have worked together to improve existing systems and develop new ones (Figure 2). In this Review, we provide a chronological overview of the intertwined development of microfluidics droplet-based compartmentalization methods and in vivo directed evolution of enzymes
Summary
Natures privileged catalysts, were optimized for a specific biological purpose and evolved over thousands of generations by natural selection. One of the main requirements in directed evolution is linking the activity of a target enzyme (i.e. the phenotype) to its genetic information (i.e. the genotype), which is essential for screening, selection and evolution campaigns.[21,22] To address this challenge, different strategies, such as compartmentalizing the enzymatic reaction within/on cells or immobilizing fluorescent products on the cell surface,. In the last twenty years, engineering and biochemistry research groups have worked together to improve existing systems and develop new ones (Figure 2) In this Review, we provide a chronological overview of the intertwined development of microfluidics droplet-based compartmentalization methods and in vivo directed evolution of enzymes
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