Explore the words cloud of the PicoCB project. It provides you a very rough idea of what is the project "PicoCB" about.
The following table provides information about the project.
THE CHANCELLOR MASTERS AND SCHOLARSOF THE UNIVERSITY OF CAMBRIDGE
|Coordinator Country||United Kingdom [UK]|
|Total cost||2˙308˙221 €|
|EC max contribution||2˙308˙221 € (100%)|
1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
|Duration (year-month-day)||from 2016-10-01 to 2021-09-30|
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|1||THE CHANCELLOR MASTERS AND SCHOLARSOF THE UNIVERSITY OF CAMBRIDGE||UK (CAMBRIDGE)||coordinator||2˙308˙221.00|
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|
Tomas Buryska, Michal Vasina, Fabrice Gielen, Pavel Vanacek, Liisa van Vliet, Jan Jezek, Zdenek Pilat, Pavel Zemanek, JiÅ™Ã DamborskÃ½, Florian Hollfelder, ZbynÄ›k 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
Hans Kleineâ€BrÃ¼ggeney, Liisa D. van Vliet, Carla Mulas, Fabrice Gielen, Chibeza C. Agley, JosÃ© C. R. Silva, Austin Smith, Kevin Chalut, Florian Hollfelder
Longâ€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
Bert van Loo, Christopher D. Bayer, Gerhard Fischer, Stefanie Jonas, Eugene Valkov, Mark F. Mohamed, Anastassia Vorobieva, Celine Dutruel, Marko HyvÃ¶nen, 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|
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
Fabrice Gielen, Raphaelle Hours, Stephane Emond, Martin Fischlechner, Ursula Schell, Florian Hollfelder
Ultrahigh-throughputâ€“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|
Tze Han Sum, Tze Jing Sum, SÃºil 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 Î² (1â€“42)
published pages: 4554-4570, ISSN: 1477-0520, DOI: 10.1039/C7OB00804J
|Organic & Biomolecular Chemistry 15/21||2020-01-22|
Fabrice Gielen, Maren Butz, Eric J. Rees, Miklos Erdelyi, Tommaso Moschetti, Marko HyvÃ¶nen, 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|
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|
Bert van Loo, Markus Schober, Eugene Valkov, Magdalena Heberlein, Erich Bornberg-Bauer, Kurt Faber, Marko HyvÃ¶nen, 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|
Charlotte M. Miton, Stefanie Jonas, Gerhard Fischer, Fernanda Duarte, Mark F. Mohamed, Bert van Loo, BÃ¡lint Kintses, Shina C. L. Kamerlin, Nobuhiko Tokuriki, Marko HyvÃ¶nen, 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|
Christopher D. Bayer, Bert vanâ€…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
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