Bioelectrochemical probing of intracellular redox processes in living yeast cells--application of redox polymer wiring in a microfluidic environment.
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.
Department of Micro- and Nanotechnology, Technical University of Denmark, Produktionstorvet 423, 2800 Kgs. Lyngby, Denmark. arto.heiskanen@nanotech.dtu.dk
Arto
A
Heiskanen
Vasile
V
Coman
Natalie
N
Kostesha
David
D
Sabourin
Nick
N
Haslett
Keith
K
Baronian
Lo
L
Gorton
Martin
M
Dufva
Jenny
J
Emnéus
D018914:Cells, Immobilized / Q000737:chemistry / Q000378:metabolism; D055664:Electrochemical Techniques / Q000295:instrumentation; D004867:Equipment Design; D005632:Fructose / Q000378:metabolism; D017353:Gene Deletion; D005947:Glucose / Q000378:metabolism; D008839:Microelectrodes; D044085:Microfluidics / Q000295:instrumentation*; D009992:Osmium / Q000737:chemistry; D010084:Oxidation-Reduction; D011108:Polymers / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D024483:Vitamin K 3 / Q000737:chemistry
D011108:Polymers; D009992:Osmium; D005632:Fructose; D024483:Vitamin K 3; D005947:Glucose