L-Shuffling™ – directed evolution

The term “gene shuffling” covers a group of directed evolution technologies that allow to create new products through random genetic recombination. These protein engineering technologies mimic a strongly accelerated natural evolution. Parent-genes are crossed amongst themselves in a random fashion in order to create a new generation of genes. This pool of new genes contains interesting combinations that code for improved proteins. These new combinations are selected and the recombination/selection cycle can be repeated as many times as necessary. Proteus’s L-shufflingTM technology is particularly powerful since it is compatible with very long sequences.

l-shuffling-scheme
l-shuffling-working

EvosightTM

EvosightTM is a method that rationalizes random mutagenesis. It is a rational approach that first explores the “protein plasticity” (ability of the protein to accept mutations with a limited loss of activity) of the enzyme, which enables us to estimate (i) the “optimal mutation load” (number of mutations introduced per gene that gives the highest frequency of improved variants) and (ii) the “ad minima size sample” (minimal number of clones to be screened) that can be used to rapidly improve this enzyme.

evosight-schemes
Chodorge M, Fourage L, Ullmann C., Duvivier V., Masson J.M., Lefèvre F. Rational Strategies for Directed Evolution of Biocatalysts – Application to
Candida antarctica lipase B (CALB) Adv. Synth. Catal.; 347:1022-1026 2005

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