Bioelectrochemical probing of intracellular redox processes in living yeast cells—application of redox polymer wiring in a microfluidic environment

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• 作者：Arto Heiskanen (1)
  Vasile Coman (1) (4)
  Natalie Kostesha (1)
  David Sabourin (1)
  Nick Haslett (2) (5)
  Keith Baronian (2)
  Lo Gorton (3)
  Martin Dufva (1)
  Jenny Emnéus (1)
  
• 关键词：Cellular redox activity ; Microbial bioelectrochemistry ; Osmium redox polymer ; Double mediator system ; Saccharomyces cerevisiae ; Microfluidic system
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2013
• 出版时间：April 2013
• 年：2013
• 卷：405
• 期：11
• 页码：3847-3858
• 全文大小：601KB
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• 作者单位：Arto Heiskanen (1)
  Vasile Coman (1) (4)
  Natalie Kostesha (1)
  David Sabourin (1)
  Nick Haslett (2) (5)
  Keith Baronian (2)
  Lo Gorton (3)
  Martin Dufva (1)
  Jenny Emnéus (1)
  
  1. Department of Micro- and Nanotechnology, Technical University of Denmark, Produktionstorvet 423, 2800, Kgs. Lyngby, Denmark
  4. Faculty of Chemistry and Chemical Engineering, Babe?-Bolyai University, Cluj-Napoca, Romania
  2. School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand
  5. Agriculture and Life Sciences Division, Lincoln University, Lincoln, Canterbury, New Zealand
  3. Department of Biochemistry and Structural Biology, Lund University, P.O. Box 124, 22100, Lund, Sweden
  
• ISSN：1618-2650

文摘

Conventionally, microbial bioelectrochemical assays have been conducted using immobilized cells on an electrode that is placed in an electrochemical batch cell. In this paper, we describe a developed microfluidic platform with integrated microelectrode arrays for automated bioelectrochemical assays utilizing a new double mediator system to map redox metabolism and screen for genetic modifications in Saccharomyces cerevisiae cells. The function of this new double mediator system based on menadione and osmium redox polymer (PVI-Os) is demonstrated. “Wiring-of S. cerevisiae cells using PVI-Os shows a significant improvement of bioelectrochemical monitoring in a microfluidic environment and functions as an effective immobilization matrix for cells that are not strongly adherent. The function of the developed microfluidic platform is demonstrated using two strains of S. cerevisiae, ENY.WA and its deletion mutant EBY44, which lacks the enzyme phosphoglucose isomerase. The cellular responses to introduced glucose and fructose were recorded for the two S. cerevisiae strains, and the obtained results are compared with previously published work when using an electrochemical batch cell, indicating that microfluidic bioelectrochemical assays employing the menadione–PVI-Os double mediator system provides an effective means to conduct automated microbial assays. Figure Microfluidic platform for bioelectrochemical assays using osmium redox polymer “wired-living yeast cells