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Real-time detection of cofactor availability in genetically modified living Saccharomyces cerevisiae cells - Simultaneous probing of different geno- and phenotypes.

Kostesha, Natalie LU ; Heiskanen, Arto LU ; Spegel, Christer LU ; Hahn-Hägerdal, Bärbel LU ; Gorwa-Grauslund, Marie-Francoise LU and Emnéus, Jenny LU (2009) In Bioelectrochemistry 76. p.180-188
Abstract
This work describes a mediated amperometric method for simultaneous real-time probing of the NAD(P)H availability in two different phenotypes, fermentative and respiratory, of the phosphoglucose isomerase deletion mutant strain of S. cerevisiae, EBY44 [ENY.WA-1A pgi1-1D::URA3], and its parental strain, ENY.WA-1A. The developed method is based on multichannel detection using microelectrode arrays. Its versatility was demonstrated by using four microelectrode arrays for simultaneously monitoring the NAD(P)H availability of both geno- and phenotypes under the influence of two different carbon sources, glucose and fructose, as well as the cytosolic and mitochondrial inhibitor and uncoupler, dicoumarol. The obtained results indicate that the... (More)
This work describes a mediated amperometric method for simultaneous real-time probing of the NAD(P)H availability in two different phenotypes, fermentative and respiratory, of the phosphoglucose isomerase deletion mutant strain of S. cerevisiae, EBY44 [ENY.WA-1A pgi1-1D::URA3], and its parental strain, ENY.WA-1A. The developed method is based on multichannel detection using microelectrode arrays. Its versatility was demonstrated by using four microelectrode arrays for simultaneously monitoring the NAD(P)H availability of both geno- and phenotypes under the influence of two different carbon sources, glucose and fructose, as well as the cytosolic and mitochondrial inhibitor and uncoupler, dicoumarol. The obtained results indicate that the method is capable of accurately and reproducibly (overall relative standard error of mean 3.2%) mapping the real-time responses of the cells with different genotype-phenotype combinations. The ENY.WA cells showed the same response to glucose and fructose when dicoumarol was used; fermentative cells indicated the presence of cytosolic inhibition and respiratory cells a net effect of mitochondrial uncoupling. EBY44 cells showed cytosolic inhibition with the exception of respiratory cells when fructose was used as carbon source. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Bioelectrochemistry
volume
76
pages
180 - 188
publisher
Elsevier
external identifiers
  • wos:000270014400028
  • pmid:19394900
  • scopus:68549115384
ISSN
1878-562X
DOI
10.1016/j.bioelechem.2009.02.015
language
English
LU publication?
yes
id
d9967a5d-8945-464d-ae1a-42d09e15a44e (old id 1391828)
date added to LUP
2009-05-12 10:04:44
date last changed
2017-11-12 03:28:42
@article{d9967a5d-8945-464d-ae1a-42d09e15a44e,
  abstract     = {This work describes a mediated amperometric method for simultaneous real-time probing of the NAD(P)H availability in two different phenotypes, fermentative and respiratory, of the phosphoglucose isomerase deletion mutant strain of S. cerevisiae, EBY44 [ENY.WA-1A pgi1-1D::URA3], and its parental strain, ENY.WA-1A. The developed method is based on multichannel detection using microelectrode arrays. Its versatility was demonstrated by using four microelectrode arrays for simultaneously monitoring the NAD(P)H availability of both geno- and phenotypes under the influence of two different carbon sources, glucose and fructose, as well as the cytosolic and mitochondrial inhibitor and uncoupler, dicoumarol. The obtained results indicate that the method is capable of accurately and reproducibly (overall relative standard error of mean 3.2%) mapping the real-time responses of the cells with different genotype-phenotype combinations. The ENY.WA cells showed the same response to glucose and fructose when dicoumarol was used; fermentative cells indicated the presence of cytosolic inhibition and respiratory cells a net effect of mitochondrial uncoupling. EBY44 cells showed cytosolic inhibition with the exception of respiratory cells when fructose was used as carbon source.},
  author       = {Kostesha, Natalie and Heiskanen, Arto and Spegel, Christer and Hahn-Hägerdal, Bärbel and Gorwa-Grauslund, Marie-Francoise and Emnéus, Jenny},
  issn         = {1878-562X},
  language     = {eng},
  pages        = {180--188},
  publisher    = {Elsevier},
  series       = {Bioelectrochemistry},
  title        = {Real-time detection of cofactor availability in genetically modified living Saccharomyces cerevisiae cells - Simultaneous probing of different geno- and phenotypes.},
  url          = {http://dx.doi.org/10.1016/j.bioelechem.2009.02.015},
  volume       = {76},
  year         = {2009},
}