Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022:2397:19-32.
doi: 10.1007/978-1-0716-1826-4_2.

Ultrahigh-Throughput Screening of Metagenomic Libraries Using Droplet Microfluidics

Davide Agostino Cecchini #  1 , Mercedes Sánchez-Costa #  1 , Alejandro H Orrego #  1 , Jesús Fernández-Lucas  2 , Aurelio Hidalgo  3
Affiliations

Ultrahigh-Throughput Screening of Metagenomic Libraries Using Droplet Microfluidics

Davide Agostino Cecchini et al. Methods Mol Biol. 2022.

Abstract

Droplet microfluidics enables the ultrahigh-throughput screening of the natural or man-made genetic diversity for industrial enzymes, with reduced reagent consumption and lower costs than conventional robotic alternatives. Here we describe an example of metagenomic screening for nucleoside 2'-deoxyribosyl transferases using FACS as a more widespread and accessible alternative than microfluidic on-chip sorters. This protocol can be easily adapted to directed evolution libraries by replacing the library construction steps and to other enzyme activities, e.g., oxidases, by replacing the proposed coupled assay.

Keywords: Enzyme screening; Functional metagenomics; Microfluidics; Nucleoside 2′-deoxyribosyl transferases.

PubMed Disclaimer

References

    1. Sheldon RA, Woodley JM (2018) Role of biocatalysis in sustainable chemistry. Chem Rev 118:801–838 - DOI
    1. May O (2019) Industrial enzyme applications—overview and historic perspective. In: Industrial enzyme applications. Wiley-VCH, Weinheim, pp 1–24. https://doi.org/10.1002/9783527813780.ch1_1 - DOI
    1. Ufarté L, Potocki-Veronese G, Laville É (2015) Discovery of new protein families and functions: new challenges in functional metagenomics for biotechnologies and microbial ecology. Front Microbiol. https://doi.org/10.3389/fmicb.2015.00563
    1. Truppo MD (2017) Biocatalysis in the pharmaceutical industry: the need for speed. ACS Med Chem Lett 8:476–480 - DOI
    1. Agresti JJ, Antipov E, Abate AR et al (2010) Ultrahigh-throughput screening in drop-based microfluidics for directed evolution. Proc Natl Acad Sci U S A 107:4004–4009 - DOI

Publication types

LinkOut - more resources