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Directed evolution of enzymes using microfluidic chips
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SYSNO ASEP 0468517 Document Type C - Proceedings Paper (int. conf.) R&D Document Type Conference Paper Title Directed evolution of enzymes using microfluidic chips Author(s) Pilát, Zdeněk (UPT-D) RID, SAI, ORCID
Ježek, Jan (UPT-D) RID, ORCID, SAI
Šmatlo, Filip (UPT-D)
Kaňka, Jan (UPT-D) RID, SAI
Zemánek, Pavel (UPT-D) RID, SAI, ORCIDNumber of authors 5 Article number 101420A Source Title 20th Slovak-Czech-Polish Optical Conference on Wave and Quantum Aspects of Contemporary Optics (Proceedings of SPIE 10142). - Bellingham : SPIE, 2016 - ISSN 0277-786X Pages s. 1-7 Number of pages 7 s. Publication form Online - E Action Slovak-Czech-Polish Optical Conference on Wave and Quantum Aspects of Contemporary Optics /20./ Event date 05.09.2016 - 09.09.2016 VEvent location Jasná Country SK - Slovakia Event type WRD Language eng - English Country US - United States Keywords bacteria ; dielectrophoresis ; engineering ; luminiscence ; proteins Subject RIV BH - Optics, Masers, Lasers R&D Projects GA16-07965S GA ČR - Czech Science Foundation (CSF) TA03010642 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) LO1212 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) ED0017/01/01 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UPT-D - RVO:68081731 UT WOS 000393152700009 EID SCOPUS 85011697938 DOI 10.1117/12.2264377 Annotation Enzymes are highly versatile and ubiquitous biological catalysts. They can greatly accelerate large variety of reactions, while ensuring appropriate catalytic activity and high selectivity. These properties make enzymes attractive biocatalysts for a wide range of industrial and biomedical applications. Over the last two decades, directed evolution of enzymes has transformed the field of protein engineering. We have devised microfluidic systems for directed evolution of haloalkane dehalogenases in emulsion droplets. In such a device, individual bacterial cells producing mutated variants of the same enzyme are encapsulated in microdroplets and supplied with a substrate. The conversion of a substrate by the enzyme produced by a single bacterium changes the pH in the droplet which is signalized by pH dependent fluorescence probe. The droplets with the highest enzymatic activity can be separated directly on the chip by dielectrophoresis and the resultant cell lineage can be used for enzyme production or for further rounds of directed evolution. This platform is applicable for fast screening of large libraries in directed evolution experiments requiring mutagenesis at multiple sites of a protein structure. Workplace Institute of Scientific Instruments Contact Martina Šillerová, sillerova@ISIBrno.Cz, Tel.: 541 514 178 Year of Publishing 2017 Electronic address http://proceedings.spiedigitallibrary.org/proceeding.aspx?articleid=2595346
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