Jump to main content
Jump to site search
Access to RSC content Close the message box

Continue to access RSC content when you are not at your institution. Follow our step-by-step guide.

Issue 1, 2020
Previous Article Next Article

Advances in ultrahigh-throughput screening for directed enzyme evolution

Author affiliations


Enzymes are versatile catalysts and their synthetic potential has been recognized for a long time. In order to exploit their full potential, enzymes often need to be re-engineered or optimized for a given application. (Semi-) rational design has emerged as a powerful means to engineer proteins, but requires detailed knowledge about structure function relationships. In turn, directed evolution methodologies, which consist of iterative rounds of diversity generation and screening, can improve an enzyme's properties with virtually no structural knowledge. Current diversity generation methods grant us access to a vast sequence space (libraries of >1012 enzyme variants) that may hide yet unexplored catalytic activities and selectivity. However, the time investment for conventional agar plate or microtiter plate-based screening assays represents a major bottleneck in directed evolution and limits the improvements that are obtainable in reasonable time. Ultrahigh-throughput screening (uHTS) methods dramatically increase the number of screening events per time, which is crucial to speed up biocatalyst design, and to widen our knowledge about sequence function relationships. In this review, we summarize recent advances in uHTS for directed enzyme evolution. We shed light on the importance of compartmentalization to preserve the essential link between genotype and phenotype and discuss how cells and biomimetic compartments can be applied to serve this function. Finally, we discuss how uHTS can inspire novel functional metagenomics approaches to identify natural biocatalysts for novel chemical transformations.

Graphical abstract: Advances in ultrahigh-throughput screening for directed enzyme evolution

Back to tab navigation

Article information

12 Jul 2019
First published
09 Dec 2019

Chem. Soc. Rev., 2020,49, 233-262
Article type
Review Article

Advances in ultrahigh-throughput screening for directed enzyme evolution

U. Markel, K. D. Essani, V. Besirlioglu, J. Schiffels, W. R. Streit and U. Schwaneberg, Chem. Soc. Rev., 2020, 49, 233
DOI: 10.1039/C8CS00981C

Social activity

Search articles by author