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Evolution of enzyme catalysts caged in biomimetic gel-shell beads

Evolution of enzyme catalysts caged in biomimetic gel-shell beads
Evolution of enzyme catalysts caged in biomimetic gel-shell beads
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 >107 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.
1755-4330
791-796
Fischlechner, Martin
b3930129-0775-4c05-81c7-475934df97ee
Schaerli, Yolanda
dad6dfa3-2474-49a6-a03d-c6282e857bb7
Mohamed, Mark F.
8b6d6351-9403-41ec-aca5-2273974ed982
Patil, Santosh
a74a0315-b118-4104-bcdd-2c2e8d3b2aae
Abell, Chris
9b071a5d-04c5-464e-8d19-e37e41d1a046
Hollfelder, Florian
a9f01280-f05d-4057-b5d7-85eac249e477
Fischlechner, Martin
b3930129-0775-4c05-81c7-475934df97ee
Schaerli, Yolanda
dad6dfa3-2474-49a6-a03d-c6282e857bb7
Mohamed, Mark F.
8b6d6351-9403-41ec-aca5-2273974ed982
Patil, Santosh
a74a0315-b118-4104-bcdd-2c2e8d3b2aae
Abell, Chris
9b071a5d-04c5-464e-8d19-e37e41d1a046
Hollfelder, Florian
a9f01280-f05d-4057-b5d7-85eac249e477

Fischlechner, Martin, Schaerli, Yolanda, Mohamed, Mark F., Patil, Santosh, Abell, Chris and Hollfelder, Florian (2014) Evolution of enzyme catalysts caged in biomimetic gel-shell beads. Nature Chemistry, 6 (9), 791-796. (doi:10.1038/nchem.1996).

Record type: Article

Abstract

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 >107 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.

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e-pub ahead of print date: 20 July 2014
Published date: 20 July 2014
Organisations: Chemistry, Faculty of Natural and Environmental Sciences

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Local EPrints ID: 369249
URI: http://eprints.soton.ac.uk/id/eprint/369249
ISSN: 1755-4330
PURE UUID: 1771933d-19f6-47af-a959-a5c5e8fbd722

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Date deposited: 30 Sep 2014 12:28
Last modified: 16 Dec 2019 20:23

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Contributors

Author: Martin Fischlechner
Author: Yolanda Schaerli
Author: Mark F. Mohamed
Author: Santosh Patil
Author: Chris Abell
Author: Florian Hollfelder

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