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Rapid assessment of Oenococcus oeni activity by measuring intracellular pH and membrane potential by flow cytometry, and its application to the more effective control of malolactic fermentation.
Int J Food Microbiol 2015; 193:139-46IJ

Abstract

The aim of this study is to highlight the changes in the physiological cellular state of Oenococcus oeni during malolactic fermentation (MLF), and to use its cellular parameters to improve existing knowledge of O. oeni behaviour and to more effectively control the performance of the bacteria during MLF in wine. To do this, measurements of intracellular pH, transmembrane potential and vitality were performed using flow cytometry with different fluorescent probes: CFDA-SE and CDCF, DiBAC and CFDA, respectively. The kinetics of the cellular changes in these parameters were determined during MLF in FT80 synthetic medium and in white wine, as were the kinetics of malic acid consumption. pHin measurement throughout the entire growth shows that the pH was equal to the pH of the culture medium during the early stage, increased to pH6 in the exponential phase, and then decreased to equilibrate with the pH of the medium in the late stationary phase. Membrane potential increased in early MLF and then decreased. The decrease in pHin and membrane potential occurred when all of the malic acid was consumed. Finally, we showed that the higher the ΔpH (pHin-pHex) in O. oeni cells was, the shorter the lag phase of the MLF was. To better manage the initiation of MLF in wines, the physiological state of O. oeni cells must be taken into account. These results allow us to understand the sometimes random initiation of MLF in wines inoculated with O. oeni and to suggest ways to improve this control.

Authors+Show Affiliations

AgroParisTech, INRA, UMR 782 Génie et Microbiologie des Procédés Alimentaires, Grignon, France. Electronic address: marielle.bouix@agroparistech.fr.AgroParisTech, INRA, UMR 782 Génie et Microbiologie des Procédés Alimentaires, Grignon, France.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

25462933

Citation

Bouix, M, and S Ghorbal. "Rapid Assessment of Oenococcus Oeni Activity By Measuring Intracellular pH and Membrane Potential By Flow Cytometry, and Its Application to the More Effective Control of Malolactic Fermentation." International Journal of Food Microbiology, vol. 193, 2015, pp. 139-46.
Bouix M, Ghorbal S. Rapid assessment of Oenococcus oeni activity by measuring intracellular pH and membrane potential by flow cytometry, and its application to the more effective control of malolactic fermentation. Int J Food Microbiol. 2015;193:139-46.
Bouix, M., & Ghorbal, S. (2015). Rapid assessment of Oenococcus oeni activity by measuring intracellular pH and membrane potential by flow cytometry, and its application to the more effective control of malolactic fermentation. International Journal of Food Microbiology, 193, pp. 139-46. doi:10.1016/j.ijfoodmicro.2014.10.019.
Bouix M, Ghorbal S. Rapid Assessment of Oenococcus Oeni Activity By Measuring Intracellular pH and Membrane Potential By Flow Cytometry, and Its Application to the More Effective Control of Malolactic Fermentation. Int J Food Microbiol. 2015 Jan 16;193:139-46. PubMed PMID: 25462933.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid assessment of Oenococcus oeni activity by measuring intracellular pH and membrane potential by flow cytometry, and its application to the more effective control of malolactic fermentation. AU - Bouix,M, AU - Ghorbal,S, Y1 - 2014/10/22/ PY - 2014/08/31/received PY - 2014/10/03/revised PY - 2014/10/17/accepted PY - 2014/12/3/entrez PY - 2014/12/3/pubmed PY - 2015/3/19/medline KW - Activity KW - Flow cytometry KW - Malolactic fermentation KW - Oenococcus oeni KW - Physiology SP - 139 EP - 46 JF - International journal of food microbiology JO - Int. J. Food Microbiol. VL - 193 N2 - The aim of this study is to highlight the changes in the physiological cellular state of Oenococcus oeni during malolactic fermentation (MLF), and to use its cellular parameters to improve existing knowledge of O. oeni behaviour and to more effectively control the performance of the bacteria during MLF in wine. To do this, measurements of intracellular pH, transmembrane potential and vitality were performed using flow cytometry with different fluorescent probes: CFDA-SE and CDCF, DiBAC and CFDA, respectively. The kinetics of the cellular changes in these parameters were determined during MLF in FT80 synthetic medium and in white wine, as were the kinetics of malic acid consumption. pHin measurement throughout the entire growth shows that the pH was equal to the pH of the culture medium during the early stage, increased to pH6 in the exponential phase, and then decreased to equilibrate with the pH of the medium in the late stationary phase. Membrane potential increased in early MLF and then decreased. The decrease in pHin and membrane potential occurred when all of the malic acid was consumed. Finally, we showed that the higher the ΔpH (pHin-pHex) in O. oeni cells was, the shorter the lag phase of the MLF was. To better manage the initiation of MLF in wines, the physiological state of O. oeni cells must be taken into account. These results allow us to understand the sometimes random initiation of MLF in wines inoculated with O. oeni and to suggest ways to improve this control. SN - 1879-3460 UR - https://www.unboundmedicine.com/medline/citation/25462933/Rapid_assessment_of_Oenococcus_oeni_activity_by_measuring_intracellular_pH_and_membrane_potential_by_flow_cytometry_and_its_application_to_the_more_effective_control_of_malolactic_fermentation_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-1605(14)00523-6 DB - PRIME DP - Unbound Medicine ER -