Synchrotron UV fluorescence microscopy for determining membrane fluidity modification of single bacteria with temperatures

Passot, Stéphanie; Jamme, Frédéric; Réfrégiers, Matthieu; Gautier, Julie; Cenard, Stéphanie; Fonseca, Fernanda

doi:10.3233/bsi-140062

Cited by 7 publications

(7 citation statements)

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“…Culture medium promoting the synthesis of UFA (increase UFA/SFA ratio), CFA, and long chain FAs made it possible to improve the LAB resistance to freezing and freezedrying processes. These membrane composition modifications seem to be correlated with an increase of membrane fluidity, in particular at low temperatures (Passot et al 2014;Gautier et al 2013). An increase of membrane fluidity can be expected when increasing CFA content due to its decreasing effect on the lipid phase transition temperature (Perly et al 1985) and from recent molecular dynamic simulations of model membranes containing CFA (Poger and Mark 2015).…”

Section: Composition Of Culture Medium and Growth In Hyperosmotic Conmentioning

confidence: 92%

“…By replacing fluorescence spectroscopy with fluorescence microscopy to measure the emitted fluorescence of the probe, it is possible to obtain subcellular mapping of membrane fluidity and to detect any membrane heterogeneity. For instance, Passot et al (2014) evidenced the formation of rigid domains within the membrane of Lb. bulgaricus cells when submitting to cold stress.…”

Section: Fluorescence Anisotropymentioning

confidence: 99%

“…Following cooling, LAB membrane fluidity decreases with the transition from the disordered liquid crystalline (Lα) to the ordered gel phase (Lβ) of the lipid bilayer. Gautier et al (2013) and Passot et al (2014) studied the evolution of membrane properties of two populations of Lb. bulgaricus CFL1 exhibiting different freezing resistance following cooling by FTIR spectroscopy and anisotropy of fluorescence (Fig.…”

Section: Modification Of Membrane Properties Following Stabilization mentioning

confidence: 99%

“…Existence of lipid domains within bacterial membrane Domains of specific lipid composition have recently been evidenced within bacterial membranes and the characterization of mechanisms underlying the local enrichment of PLs has become an active research area (Romantsov et al 2009;Passot et al 2014;Lin and Weibel 2016). Membrane poles and septa of bacilli including Gram-positive bacteria were reported to be enriched in anionic phospholipids, especially cardiolipin (CL) (Kawai et al 2004;López 2006;Bernal et al 2007;Seydlová et al 2013).…”

Section: Future Prospects For Research On Lab Membranesmentioning

confidence: 99%

“…The fermentation conditions and the resulting membrane lipid composition are commonly related to the resistance of LAB to stabilization processes. Even it is well admitted that membrane fluidity is governed by the fatty acid composition of the membrane, few works report direct assessment of membrane fluidity after fermentation (Velly et al 2015;Bouix and Ghorbal 2017) and following stabilization process (Schwab et al 2007;Passot et al 2014;Meneghel et al 2017b).…”

Section: Introductionmentioning

confidence: 99%

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Factors influencing the membrane fluidity and the impact on production of lactic acid bacteria starters

Fonseca

Pénicaud

Tymczyszyn

et al. 2019

Appl Microbiol Biotechnol

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Production of lactic acid bacteria starters for manufacturing food, probiotic, and chemical products requires the application of successive steps: fermentation, concentration, stabilization, and storage. Despite process optimization, losses of bacterial viability and functional activities are observed after stabilization and storage steps due to cell exposure to environmental stresses (thermal, osmotic, mechanical, and oxidative). Bacterial membrane is the primary target for injury and its damage is highly dependent on its physical properties and lipid organization. Membrane fluidity is a key property for maintaining cell functionality, and depends on lipid composition and cell environment. Extensive evidence has been reported on changes in membrane fatty acyl chains when modifying fermentation conditions. However, a deep characterization of membrane physical properties and their evolution following production processes is scarcely reported. Therefore, the aims of this mini-review are (i) to define the membrane fluidity and the methods used to assess it and (ii) to summarize the effect of environmental conditions on membrane fluidity and the resulting impact on the resistance of lactic acid bacteria to the stabilization processes. This will make it possible to highlight existing gaps of knowledge and opens up novel approaches for future investigations.

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Section: Composition Of Culture Medium and Growth In Hyperosmotic Conmentioning

confidence: 92%

Section: Fluorescence Anisotropymentioning

confidence: 99%

Section: Modification Of Membrane Properties Following Stabilization mentioning

confidence: 99%

Section: Future Prospects For Research On Lab Membranesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Factors influencing the membrane fluidity and the impact on production of lactic acid bacteria starters

Fonseca

Pénicaud

Tymczyszyn

et al. 2019

Appl Microbiol Biotechnol

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View full text Add to dashboard Cite

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Insights into lactic acid bacteria cryoresistance using FTIR microspectroscopy

et al. 2021

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Freezing is widely used for bacterial cell preservation. However, resistance to freezing can greatly vary depending on bacterial species or growth conditions. Our study aims at identifying cellular markers of cryoresistance based on the comparison of three lactic acid bacteria (LAB) exhibiting different tolerance to freezing: Carnobacterium maltaromaticum CNCM I-3298, Lactobacillus delbrueckii subsp.bulgaricus ATCC 11842, and Lactobacillus delbrueckii subsp. bulgaricus CFL1.A thorough characterization of their cytoplasmic membrane properties was carried out by measuring their fatty acid composition, membrane fluidity, and lipid phase transition upon cooling from 50 °C to -50 °C. Vitrification temperatures of the intra-and extra-cellular compartments were also quantified by differential scanning calorimetry. Additionally, the cell biochemical characterization was carried out using a recently developed Fourier transform infrared (FTIR) micro-spectroscopic approach allowing the analysis of live bacteria in an aqueous environment.The multivariate analysis of the FTIR spectra of fresh and thawed cells enabled the discrimination of the three bacteria according to their lipid, protein, and cell wall peptidoglycan components. It also revealed freezing-induced modifications of these three cellular components and an increase in bacteria heterogeneity for the two strains of L. bulgaricus, the freeze-sensitive bacteria. No cellular damage was observed for C. maltaromaticum, the freeze-resistant bacteria. Comparison of the results obtained from the different analytical methods confirmed previously reported cryoresistance markers and suggested new ones, such as changes in the absorbance of specific infrared spectral bands. FTIR microspectroscopy could be used as a rapid and non-invasive technique to evaluate the freeze-sensitivity of LAB.

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Biophysical characterization of the Lactobacillus delbrueckii subsp. bulgaricus membrane during cold and osmotic stress and its relevance for cryopreservation

Meneghel

Passot

Dupont

et al. 2016

Appl Microbiol Biotechnol

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Freezing lactic acid bacteria often leads to cell death and loss of technological properties. Our objective was to provide an in-depth characterization of the biophysical properties of the Lactobacillus delbrueckii subsp. bulgaricus membrane in relation to its freeze resistance. Freezing was represented as a combination of cold and osmotic stress. This work investigated the relative incidence of increasing sucrose concentrations coupled or not with subzero temperatures without ice nucleation on the biological and biophysical responses of two strains with different membrane fatty acid compositions and freeze resistances. Following exposure of bacterial cells to the highest sucrose concentration, the sensitive strain exhibited a survival rate of less than 10 % and 5 h of acidifying activity loss. Similar biological activity losses were observed upon freeze-thawing and after osmotic treatment for each strain thus highlighting osmotic stress as the main source of cryoinjury. The direct measurement of membrane fluidity by fluorescence anisotropy was linked to membrane lipid organization characterized by FTIR spectroscopy. Both approaches made it possible to investigate the specific contributions of the membrane core and the bilayer external surface to cell degradation caused by cold and osmotic stress. Cold-induced membrane rigidification had no significant implication on bacterial freeze-thaw resistance. Interactions between extracellular sucrose and membrane phospholipid headgroups under osmotic stress were also observed. Such interactions were more evident in the sensitive strain and when increasing sucrose concentration, thus suggesting membrane permeabilization. The relevance of biophysical properties for elucidating mechanisms of cryoinjury and cryoprotection is discussed.

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Synchrotron UV fluorescence microscopy for determining membrane fluidity modification of single bacteria with temperatures

Cited by 7 publications

References 0 publications

Factors influencing the membrane fluidity and the impact on production of lactic acid bacteria starters

Factors influencing the membrane fluidity and the impact on production of lactic acid bacteria starters

Insights into lactic acid bacteria cryoresistance using FTIR microspectroscopy

Biophysical characterization of the Lactobacillus delbrueckii subsp. bulgaricus membrane during cold and osmotic stress and its relevance for cryopreservation

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