Literature DB >> 32896286

Ultrahigh throughput screening for enzyme function in droplets.

Stefanie Neun1, Paul J Zurek1, Tomasz S Kaminski1, Florian Hollfelder2.   

Abstract

Water-in-oil droplets, made and handled in microfluidic devices, provide a new experimental format, in which ultrahigh throughput experiments can be conducted faster and with minimal reagent consumption. An increasing number of studies have emerged that applied this approach to directed evolution and metagenomic screening of enzyme catalysts. Here, we review the considerations necessary to implement robust workflows, based on choices of device design, detection modes, emulsion formulations and substrates, and scope out which enzyme classes have become amenable to droplet screening.
© 2020 Elsevier Inc. All rights reserved.

Entities:  

Keywords:  Catalytic promiscuity; Directed evolution; Functional metagenomics; Hydrolase; In vitro compartmentalization; Machine learning; Microdroplets; Next generation sequencing; Oxidoreductase; Ultrahigh throughput screening

Mesh:

Substances:

Year:  2020        PMID: 32896286     DOI: 10.1016/bs.mie.2020.06.002

Source DB:  PubMed          Journal:  Methods Enzymol        ISSN: 0076-6879            Impact factor:   1.600


  6 in total

1.  Functional metagenomic screening identifies an unexpected β-glucuronidase.

Authors:  Stefanie Neun; Paul Brear; Eleanor Campbell; Theodora Tryfona; Kamel El Omari; Armin Wagner; Paul Dupree; Marko Hyvönen; Florian Hollfelder
Journal:  Nat Chem Biol       Date:  2022-07-07       Impact factor: 16.174

2.  Ultrahigh-Throughput Detection of Enzymatic Alcohol Dehydrogenase Activity in Microfluidic Droplets with a Direct Fluorogenic Assay.

Authors:  Miriam Klaus; Paul Jannis Zurek; Tomasz S Kaminski; Ahir Pushpanath; Katharina Neufeld; Florian Hollfelder
Journal:  Chembiochem       Date:  2021-10-13       Impact factor: 3.461

3.  Cell-free Directed Evolution of a Protease in Microdroplets at Ultrahigh Throughput.

Authors:  Josephin M Holstein; Christian Gylstorff; Florian Hollfelder
Journal:  ACS Synth Biol       Date:  2021-01-27       Impact factor: 5.110

Review 4.  Directed Evolution in Drops: Molecular Aspects and Applications.

Authors:  Aitor Manteca; Alejandra Gadea; David Van Assche; Pauline Cossard; Mélanie Gillard-Bocquet; Thomas Beneyton; C Axel Innis; Jean-Christophe Baret
Journal:  ACS Synth Biol       Date:  2021-10-22       Impact factor: 5.110

Review 5.  Recent trends in biocatalysis.

Authors:  Dong Yi; Thomas Bayer; Christoffel P S Badenhorst; Shuke Wu; Mark Doerr; Matthias Höhne; Uwe T Bornscheuer
Journal:  Chem Soc Rev       Date:  2021-06-18       Impact factor: 60.615

6.  A Titratable Cell Lysis-on-Demand System for Droplet-Compartmentalized Ultrahigh-Throughput Screening in Functional Metagenomics and Directed Evolution.

Authors:  Che Fai Alex Wong; Liisa van Vliet; Swapnil Vilas Bhujbal; Chengzhi Guo; Marit Sletmoen; Bjørn Torger Stokke; Florian Hollfelder; Rahmi Lale
Journal:  ACS Synth Biol       Date:  2021-07-14       Impact factor: 5.110

  6 in total

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