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Enzyme engineering in biomimetic compartments.


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Authors

Colin, Pierre-Yves 
Zinchenko, Anastasia 
Hollfelder, Florian  ORCID logo  https://orcid.org/0000-0002-1367-6312

Abstract

The success of a directed evolution approach to creating custom-made enzymes relies in no small part on screening as many clones as possible. The miniaturisation of assays into pico to femtoliter compartments (emulsion droplets, vesicles or gel-shell beads) makes directed evolution campaigns practically more straightforward than current large scale industrial screening that requires liquid handling equipment and much manpower. Several recent experimental formats have established protocols to screen more than 10 million compartments per day, representing unprecedented throughput at low cost. This review introduces the emerging approaches towards making biomimetic man-made compartments that are poised to be adapted by a wider circle of researchers. In addition to cost and time saving, control of selection pressures and conditions, the quantitative readout that reports on every library members and the ability to develop strategies based on these data will increase the degrees of freedom in designing and testing strategies for directed evolution experiments.

Description

Keywords

Biomimetics, Directed Molecular Evolution, Emulsions, Enzymes, Lab-On-A-Chip Devices, Microchip Analytical Procedures, Protein Engineering

Journal Title

Curr Opin Struct Biol

Conference Name

Journal ISSN

0959-440X
1879-033X

Volume Title

33

Publisher

Elsevier BV
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/L002469/1)
Engineering and Physical Sciences Research Council (EP/H046593/1)
Our research was funded by the Biological and Biotechnological Research Council (BBSRC) and the Engineering and Physical Sciences Research Council (EPSRC). FH is an ERC Starting Investigator, PYC and AZ were supported by the EU Marie-Curie training network PhosChemRec. AZ received further support from the BBSRC and the Cambridge European Trust.