Activity of ethanol-stressed Oenococcus oeni cells: a flow cytometric approach

M.G. da Silveira, T. Abee

Research output: Contribution to journalArticleAcademicpeer-review

12 Citations (Scopus)

Abstract

Aims: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. Methods and Results: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis revealed that O. oeni cells extrude the accumulated cF upon energizing with l-malic acid. However, O. oeni cells exposed to 12% (v/v) ethanol for 1 h showed a decreased capacity for active extrusion of cF. Moreover, two subpopulations could be distinguished, one of which being able to extrude cF and the other one remaining cF fluorescent. Growing cells in the presence of 8% (v/v) ethanol resulted in robust cells that maintained the capacity to actively extrude cF after being exposed to 12% (v/v) ethanol, which in turn correlated with the high levels of ATP observed in these ethanol stressed, malolactic fermentation (MLF) performing cells. Conclusion: From our results, it becomes evident that active extrusion of cF can be used to assess malolactic activity in O. oeni. Significance and Impact of the Study: The present study provides information for the development of a rapid method to assess the malolactic activity of individual O. oeni cells performing MLF during wine production
Original languageEnglish
Pages (from-to)1690-1696
JournalJournal of Applied Microbiology
Volume106
Issue number5
DOIs
Publication statusPublished - 2009

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Oenococcus
Fluorescein
Ethanol
Fermentation
Wine
Fluorescent Dyes
Adenosine Triphosphate

Keywords

  • electrogenic malate uptake
  • lactic-acid bacteria
  • leuconostoc-oenos
  • malolactic fermentation
  • lactococcus-lactis
  • fluorescent-probe
  • membrane
  • energy
  • glucose
  • growth

Cite this

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title = "Activity of ethanol-stressed Oenococcus oeni cells: a flow cytometric approach",
abstract = "Aims: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. Methods and Results: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis revealed that O. oeni cells extrude the accumulated cF upon energizing with l-malic acid. However, O. oeni cells exposed to 12{\%} (v/v) ethanol for 1 h showed a decreased capacity for active extrusion of cF. Moreover, two subpopulations could be distinguished, one of which being able to extrude cF and the other one remaining cF fluorescent. Growing cells in the presence of 8{\%} (v/v) ethanol resulted in robust cells that maintained the capacity to actively extrude cF after being exposed to 12{\%} (v/v) ethanol, which in turn correlated with the high levels of ATP observed in these ethanol stressed, malolactic fermentation (MLF) performing cells. Conclusion: From our results, it becomes evident that active extrusion of cF can be used to assess malolactic activity in O. oeni. Significance and Impact of the Study: The present study provides information for the development of a rapid method to assess the malolactic activity of individual O. oeni cells performing MLF during wine production",
keywords = "electrogenic malate uptake, lactic-acid bacteria, leuconostoc-oenos, malolactic fermentation, lactococcus-lactis, fluorescent-probe, membrane, energy, glucose, growth",
author = "{da Silveira}, M.G. and T. Abee",
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language = "English",
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Activity of ethanol-stressed Oenococcus oeni cells: a flow cytometric approach. / da Silveira, M.G.; Abee, T.

In: Journal of Applied Microbiology, Vol. 106, No. 5, 2009, p. 1690-1696.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Activity of ethanol-stressed Oenococcus oeni cells: a flow cytometric approach

AU - da Silveira, M.G.

AU - Abee, T.

PY - 2009

Y1 - 2009

N2 - Aims: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. Methods and Results: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis revealed that O. oeni cells extrude the accumulated cF upon energizing with l-malic acid. However, O. oeni cells exposed to 12% (v/v) ethanol for 1 h showed a decreased capacity for active extrusion of cF. Moreover, two subpopulations could be distinguished, one of which being able to extrude cF and the other one remaining cF fluorescent. Growing cells in the presence of 8% (v/v) ethanol resulted in robust cells that maintained the capacity to actively extrude cF after being exposed to 12% (v/v) ethanol, which in turn correlated with the high levels of ATP observed in these ethanol stressed, malolactic fermentation (MLF) performing cells. Conclusion: From our results, it becomes evident that active extrusion of cF can be used to assess malolactic activity in O. oeni. Significance and Impact of the Study: The present study provides information for the development of a rapid method to assess the malolactic activity of individual O. oeni cells performing MLF during wine production

AB - Aims: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. Methods and Results: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis revealed that O. oeni cells extrude the accumulated cF upon energizing with l-malic acid. However, O. oeni cells exposed to 12% (v/v) ethanol for 1 h showed a decreased capacity for active extrusion of cF. Moreover, two subpopulations could be distinguished, one of which being able to extrude cF and the other one remaining cF fluorescent. Growing cells in the presence of 8% (v/v) ethanol resulted in robust cells that maintained the capacity to actively extrude cF after being exposed to 12% (v/v) ethanol, which in turn correlated with the high levels of ATP observed in these ethanol stressed, malolactic fermentation (MLF) performing cells. Conclusion: From our results, it becomes evident that active extrusion of cF can be used to assess malolactic activity in O. oeni. Significance and Impact of the Study: The present study provides information for the development of a rapid method to assess the malolactic activity of individual O. oeni cells performing MLF during wine production

KW - electrogenic malate uptake

KW - lactic-acid bacteria

KW - leuconostoc-oenos

KW - malolactic fermentation

KW - lactococcus-lactis

KW - fluorescent-probe

KW - membrane

KW - energy

KW - glucose

KW - growth

U2 - 10.1111/j.1365-2672.2008.04136.x

DO - 10.1111/j.1365-2672.2008.04136.x

M3 - Article

VL - 106

SP - 1690

EP - 1696

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

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