Evolution of enzyme catalysts caged in biomimetic gel-shell beads.

Détails

ID Serval
serval:BIB_99EBD3512773
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Evolution of enzyme catalysts caged in biomimetic gel-shell beads.
Périodique
Nature Chemistry
Auteur⸱e⸱s
Fischlechner M., Schaerli Y., Mohamed M.F., Patil S., Abell C., Hollfelder F.
ISSN
1755-4349 (Electronic)
ISSN-L
1755-4330
Statut éditorial
Publié
Date de publication
2014
Volume
6
Numéro
9
Pages
791-796
Langue
anglais
Résumé
Natural evolution relies on the improvement of biological entities by rounds of diversification and selection. In the laboratory, directed evolution has emerged as a powerful tool for the development of new and improved biomolecules, but it is limited by the enormous workload and cost of screening sufficiently large combinatorial libraries. Here we describe the production of gel-shell beads (GSBs) with the help of a microfluidic device. These hydrogel beads are surrounded with a polyelectrolyte shell that encloses an enzyme, its encoding DNA and the fluorescent reaction product. Active clones in these man-made compartments can be identified readily by fluorescence-activated sorting at rates >10(7) GSBs per hour. We use this system to perform the directed evolution of a phosphotriesterase (a bioremediation catalyst) caged in GSBs and isolate a 20-fold faster mutant in less than one hour. We thus establish a practically undemanding method for ultrahigh-throughput screening that results in functional hybrid composites endowed with evolvable protein components.

Mots-clé
Biomimetics, Catalysis, Gels, Microfluidic Analytical Techniques, Phosphoric Diester Hydrolases/chemistry
Pubmed
Web of science
Création de la notice
02/02/2017 8:04
Dernière modification de la notice
20/08/2019 15:01
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