Improvement of microtas for directed evolution of NAD(P)-dependent oxidoreductases

Haruna Goto, Yuki Kanai, Arisa Yotsui, Shota Shimokihara, Shunya Shitara, Yasuaki Einaga, Yoshinori Matsumoto, Norihisa Miki, Kei Fujiwara, Nobuhide Doi

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Here we report an improved μTAS based on electrochemical measurements and microbeads sorting for screening of NAD(P)-dependent oxidoreductases. By using this system, we measured the activity of model enzymes immobilized on microbeads by detection of NADH with a boron-doped diamond (BDD) electrode in each compartment (∼30 nL) of the microfluidic system. Then, we recovered the magnetic microbeads or the droplets containing genes of enzymes with higher activity by magnetophoretic or dielectrophoretic sorting, respectively. Our μTAS should be useful for directed evolution of various NAD(P)-dependent oxidoreductases with high activity and stability in the future.

Original languageEnglish
Title of host publication22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018
PublisherChemical and Biological Microsystems Society
Pages2247-2249
Number of pages3
ISBN (Electronic)9781510897571
Publication statusPublished - 2018 Jan 1
Event22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018 - Kaohsiung, Taiwan, Province of China
Duration: 2018 Nov 112018 Nov 15

Publication series

Name22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018
Volume4

Conference

Conference22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018
CountryTaiwan, Province of China
CityKaohsiung
Period18/11/1118/11/15

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Keywords

  • Boron-doped diamond electrode
  • Cell-free protein synthesis
  • Dielectrophoresis
  • Directed evolution
  • High-throughput screening

ASJC Scopus subject areas

  • Chemistry(all)
  • Bioengineering
  • Chemical Engineering (miscellaneous)
  • Control and Systems Engineering

Cite this

Goto, H., Kanai, Y., Yotsui, A., Shimokihara, S., Shitara, S., Einaga, Y., Matsumoto, Y., Miki, N., Fujiwara, K., & Doi, N. (2018). Improvement of microtas for directed evolution of NAD(P)-dependent oxidoreductases. In 22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018 (pp. 2247-2249). (22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018; Vol. 4). Chemical and Biological Microsystems Society.