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Probing the redox metabolism in the strictly anaerobic, extremely thermophilic, hydrogen-producing Caldicellulosiruptor saccharolyticus using amperometry

Kostesha, Natalie ; Willquist, Karin LU ; Emneus, Jenny and van Niel, Ed LU (2011) In Extremophiles 15(1). p.77-87
Abstract
Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular electron flow were observed when cells were supplied with different carbon sources. A higher electrochemical response was detected when cells were supplied with xylose than with sucrose or glucose. Moreover, using the mediated electrochemical method, it was possible to detect differences in the electron flow between cells harvested in the exponential and stationary growth phases. The electron flow of C. saccharolyticus was dependent on the NADH-... (More)
Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular electron flow were observed when cells were supplied with different carbon sources. A higher electrochemical response was detected when cells were supplied with xylose than with sucrose or glucose. Moreover, using the mediated electrochemical method, it was possible to detect differences in the electron flow between cells harvested in the exponential and stationary growth phases. The electron flow of C. saccharolyticus was dependent on the NADH- and reduced ferredoxin generation flux and the competitive behavior of cytosolic and membrane-associated oxidoreductases. Sodium oxamate was used to inhibit the NADH-dependent lactate dehydrogenase, upon which more NADH was directed to membrane-associated enzymes for ferricyanide reduction, leading to a higher electrochemical signal. The method is noninvasive and the results presented here demonstrate that this method can be used to accurately detect changes in the intracellular electron flow and to probe redox enzyme properties of a strictly anaerobic thermophile in vivo. (Less)
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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Caldicellulosiruptor saccharolyticus, Lactate regulation, Anaerobiosis, Electrochemistry, Mediated amperometry, Ferricyanide
in
Extremophiles
volume
15
issue
1
pages
77 - 87
publisher
Springer
external identifiers
  • wos:000286004100008
  • scopus:78651234029
  • pmid:21132340
ISSN
1433-4909
DOI
10.1007/s00792-010-0341-4
language
English
LU publication?
yes
id
3db34d71-8169-4b4c-97e8-b469a52e4956 (old id 1790902)
date added to LUP
2016-04-01 10:50:51
date last changed
2022-01-26 03:00:59
@article{3db34d71-8169-4b4c-97e8-b469a52e4956,
  abstract     = {{Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular electron flow were observed when cells were supplied with different carbon sources. A higher electrochemical response was detected when cells were supplied with xylose than with sucrose or glucose. Moreover, using the mediated electrochemical method, it was possible to detect differences in the electron flow between cells harvested in the exponential and stationary growth phases. The electron flow of C. saccharolyticus was dependent on the NADH- and reduced ferredoxin generation flux and the competitive behavior of cytosolic and membrane-associated oxidoreductases. Sodium oxamate was used to inhibit the NADH-dependent lactate dehydrogenase, upon which more NADH was directed to membrane-associated enzymes for ferricyanide reduction, leading to a higher electrochemical signal. The method is noninvasive and the results presented here demonstrate that this method can be used to accurately detect changes in the intracellular electron flow and to probe redox enzyme properties of a strictly anaerobic thermophile in vivo.}},
  author       = {{Kostesha, Natalie and Willquist, Karin and Emneus, Jenny and van Niel, Ed}},
  issn         = {{1433-4909}},
  keywords     = {{Caldicellulosiruptor saccharolyticus; Lactate regulation; Anaerobiosis; Electrochemistry; Mediated amperometry; Ferricyanide}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{77--87}},
  publisher    = {{Springer}},
  series       = {{Extremophiles}},
  title        = {{Probing the redox metabolism in the strictly anaerobic, extremely thermophilic, hydrogen-producing Caldicellulosiruptor saccharolyticus using amperometry}},
  url          = {{http://dx.doi.org/10.1007/s00792-010-0341-4}},
  doi          = {{10.1007/s00792-010-0341-4}},
  volume       = {{15}},
  year         = {{2011}},
}