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Exploring the Chemical Biology of Sequence Space via Picoliter Droplets

Total Cost €


EC-Contrib. €






 PicoCB project word cloud

Explore the words cloud of the PicoCB project. It provides you a very rough idea of what is the project "PicoCB" about.

metagenomic    hundred    fitness    proteins    evolution    emerged    capturing    interconversion    functional    vivo    enzyme    protein    made    libraries    data    microfluidic    vitro    capacity    quantitative    screened    superfamilies    tens    chance    mutations    lower    catalytic    screen    variants    lines    enzymes    shown    fundamental    oil    dollars    biocatalysis    gene    cheaper    guidelines    promiscuous    complement    catalysts    phenotype    screening    landscapes    unprecedented    interpretation    frequencies    millions    evolutionary    successful    indel    strategic    experiments    promiscuity    constitutes    arguably    record    microdroplets    random    directed    catalysis    compartment    specificity    khz    facilities    format    enormous    conducting    individual    multiple    environmental    signalling    mechanistic    reaction    group    costly    reactivity    industrial    water    droplet    engineering    members    rare    cumbersome    biological    link    monodisperse    formats    networks    conventional    encoding    plan    function    governing    led    characterise    genotype    collections    compartments    throughput    library    investment   

Project "PicoCB" data sheet

The following table provides information about the project.


Organization address
postcode: CB2 1TN

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country United Kingdom [UK]
 Total cost 2˙308˙221 €
 EC max contribution 2˙308˙221 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2015-AdG
 Funding Scheme ERC-ADG
 Starting year 2016
 Duration (year-month-day) from 2016-10-01   to  2021-09-30


Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 


 Project objective

Directed evolution of functional proteins has arguably emerged as an approach to protein engineering that can complement or better design-led approaches to protein function. However, as a random process, enormous numbers of variants have to be screened and selected to have a chance to identify successful catalysts. This process is costly and cumbersome: Industrial screening facilities require investment of tens to hundred millions of dollars. My group has implemented key steps towards conducting quantitative biological experiments in a much cheaper format. Screening of individual library members in monodisperse oil-in-water compartments ('microdroplets’) that are generated at kHz frequencies in microfluidic devices has been shown to be possible. The droplet compartment constitutes a link between a given phenotype and its encoding genotype, by capturing reaction product, and thus providing a unique system to screen for catalysis.In this way quantitative fitness landscapes for interconversion of members of enzyme superfamilies along the lines of catalytic promiscuity, understanding the factors governing specificity and the mechanistic interpretation of the observed evolutionary pathways can be made. We now apply this screening system of unprecedented capacity for directed evolution and metagenomic screening of enzymes in in vivo and in vitro formats. We plan to apply this system to do experiments that would not be possible with conventional, lower throughput approaches: (i) screening of metagenomic libraries for rare and promiscuous activities that characterise environmental gene collections for their reactivity and potential for applied biocatalysis; (ii) developing a fundamental understanding of and strategic guidelines for enzyme evolution based on fitness landscapes that record data on multiple, promiscuous activities in response to Indel mutations; and (iii) evolution of gene networks to build up signalling networks in vitro.


year authors and title journal last update
List of publications.
2019 Tomas Buryska, Michal Vasina, Fabrice Gielen, Pavel Vanacek, Liisa van Vliet, Jan Jezek, Zdenek Pilat, Pavel Zemanek, Jiří Damborský, Florian Hollfelder, Zbyněk Prokop
Controlled oil/water partitioning of hydrophobic substrates extends the bioanalytical applications of droplet-based microfluidics
published pages: , ISSN: 0003-2700, DOI: 10.1021/acs.analchem.9b01839
Analytical Chemistry 2020-01-22
2019 Hans Kleine‐Brüggeney, Liisa D. van Vliet, Carla Mulas, Fabrice Gielen, Chibeza C. Agley, José C. R. Silva, Austin Smith, Kevin Chalut, Florian Hollfelder
Long‐Term Perfusion Culture of Monoclonal Embryonic Stem Cells in 3D Hydrogel Beads for Continuous Optical Analysis of Differentiation
published pages: 1804576, ISSN: 1613-6810, DOI: 10.1002/smll.201804576
Small 15/5 2020-01-22
2018 Bert van Loo, Christopher D. Bayer, Gerhard Fischer, Stefanie Jonas, Eugene Valkov, Mark F. Mohamed, Anastassia Vorobieva, Celine Dutruel, Marko Hyvönen, Florian Hollfelder
Balancing Specificity and Promiscuity in Enzyme Evolution: Multidimensional Activity Transitions in the Alkaline Phosphatase Superfamily
published pages: 370-387, ISSN: 0002-7863, DOI: 10.1021/jacs.8b10290
Journal of the American Chemical Society 141/1 2020-01-22
2019 Bert van Loo, Ryan Berry, Usa Boonyuen, Mark F. Mohamed, Marko Golicnik, Alvan C. Hengge, Florian Hollfelder
Transition-State Interactions in a Promiscuous Enzyme: Sulfate and Phosphate Monoester Hydrolysis by Pseudomonas aeruginosa Arylsulfatase
published pages: 1363-1378, ISSN: 0006-2960, DOI: 10.1021/acs.biochem.8b00996
Biochemistry 58/10 2020-01-22
2016 Fabrice Gielen, Raphaelle Hours, Stephane Emond, Martin Fischlechner, Ursula Schell, Florian Hollfelder
Ultrahigh-throughput–directed enzyme evolution by absorbance-activated droplet sorting (AADS)
published pages: E7383-E7389, ISSN: 0027-8424, DOI: 10.1073/pnas.1606927113
Proceedings of the National Academy of Sciences 113/47 2020-01-22
2017 Tze Han Sum, Tze Jing Sum, Súil Collins, Warren R. J. D. Galloway, David G. Twigg, Florian Hollfelder, David R. Spring
Divergent synthesis of biflavonoids yields novel inhibitors of the aggregation of amyloid β (1–42)
published pages: 4554-4570, ISSN: 1477-0520, DOI: 10.1039/C7OB00804J
Organic & Biomolecular Chemistry 15/21 2020-01-22
2016 Fabrice Gielen, Maren Butz, Eric J. Rees, Miklos Erdelyi, Tommaso Moschetti, Marko Hyvönen, Joshua B. Edel, Clemens F. Kaminski, Florian Hollfelder
Quantitative Affinity Determination by Fluorescence Anisotropy Measurements of Individual Nanoliter Droplets
published pages: 1092-1101, ISSN: 0003-2700, DOI: 10.1021/acs.analchem.6b02528
Analytical Chemistry 89/2 2020-01-22
2016 Miriam Kaltenbach, Stephane Emond, Florian Hollfelder, Nobuhiko Tokuriki
Functional Trade-Offs in Promiscuous Enzymes Cannot Be Explained by Intrinsic Mutational Robustness of the Native Activity
published pages: e1006305, ISSN: 1553-7404, DOI: 10.1371/journal.pgen.1006305
PLOS Genetics 12/10 2020-01-22
2018 Bert van Loo, Markus Schober, Eugene Valkov, Magdalena Heberlein, Erich Bornberg-Bauer, Kurt Faber, Marko Hyvönen, Florian Hollfelder
Structural and Mechanistic Analysis of the Choline Sulfatase from Sinorhizobium melliloti : A Class I Sulfatase Specific for an Alkyl Sulfate Ester
published pages: 1004-1023, ISSN: 0022-2836, DOI: 10.1016/j.jmb.2018.02.010
Journal of Molecular Biology 430/7 2020-01-22
2018 Charlotte M. Miton, Stefanie Jonas, Gerhard Fischer, Fernanda Duarte, Mark F. Mohamed, Bert van Loo, Bálint Kintses, Shina C. L. Kamerlin, Nobuhiko Tokuriki, Marko Hyvönen, Florian Hollfelder
Evolutionary repurposing of a sulfatase: A new Michaelis complex leads to efficient transition state charge offset
published pages: E7293-E7302, ISSN: 0027-8424, DOI: 10.1073/pnas.1607817115
Proceedings of the National Academy of Sciences 115/31 2020-01-22
2017 Christopher D. Bayer, Bert van Loo, Florian Hollfelder
Specificity Effects of Amino Acid Substitutions in Promiscuous Hydrolases: Context-Dependence of Catalytic Residue Contributions to Local Fitness Landscapes in Nearby Sequence Space
published pages: 1001-1015, ISSN: 1439-4227, DOI: 10.1002/cbic.201600657
ChemBioChem 18/11 2020-01-22

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