Details

Autor(en) / Beteiligte

• Yang, Guangyu
• Withers, Stephen G.

Titel

Ultrahigh-Throughput FACS-Based Screening for Directed Enzyme Evolution

Ist Teil von

• Chembiochem : a European journal of chemical biology, 2009-11, Vol.10 (17), p.2704-2715

Ort / Verlag

Weinheim: WILEY-VCH Verlag

Erscheinungsjahr

2009

Quelle

MEDLINE

Beschreibungen/Notizen

• Directed enzyme evolution has proven to be a powerful tool for improving a range of properties of enzymes through consecutive rounds of diversification and selection. However, its success depends heavily on the efficiency of the screening strategy employed. Fluorescence‐activated cell sorting (FACS) has recently emerged as a powerful tool for screening enzyme libraries due to its high sensitivity and its ability to analyze as many as 108 mutants per day. Applications of FACS screening have allowed the isolation of enzyme variants with significantly improved activities, altered substrate specificities, or even novel functions. This review discusses FACS‐based screening for enzymatic activity and its potential application for the directed evolution of enzymes, ribozymes, and catalytic antibodies. The FACS of life: Fluorescence‐activated cell sorting (FACS) has recently emerged as a powerful tool for screening enzyme libraries. The key step in developing a FACS screening method is to establish a linkage between genotype and phenotype. In this minireview, we discuss recent advances in FACS‐based screening for enzymatic activity and especially focus on the novel approaches that couple the target enzymatic activity with a detectable fluorescent signal.