Pulsed electric field for increasing intracellular trehalose content in Lactobacillus plantarum WCFS1

Vaessen, Evelien M.J.; Besten, H.M.W. den; Patra, Tiffany; Mossevelde, N.T.M. van; Boom, R.M.; Schutyser, M.A.I.

doi:10.1016/j.ifset.2018.03.007

Cited by 28 publications

(16 citation statements)

References 27 publications

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“…For every PEF experiment a fresh stationary phase culture of Lactobacillus plantarum WCFS1 was used. L. plantarum WCFS1 was cultured in the same way as described in previous research 9 . Briefly, fresh cultures were prepared from a frozen stock culture (−80 °C) on De Man Rogosa and Sharpe (MRS) (Merck, Germany) agar (Oxoid, United Kingdom) plates.…”

Section: Methodsmentioning

confidence: 99%

“…This stationary phase culture (9.5 log CFU/mL) was centrifuged for 10 minutes at 13,500 × g at 20 °C, and the resulting pellet was washed once with washing solution and subsequently suspended in electroporation medium, containing 0.3 M trehalose (Merck, Germany). The exact compositions of the washing solution and electroporation medium can be found in previous research 9 .The final conductivity and pH of the suspended culture (9.5 log CFU/mL) in electroporation medium at 21 °C were 0.15 S/m and pH 6.8, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Loading of small molecules, such as trehalose, into cells has been shown to increase the processing robustness of mammalian cells and plant tissue 7,8 . Our previous study demonstrated that PEF treatment can also be used to increase intracellular trehalose in the model probiotic bacterium, Lactobacillus plantarum WCFS1, while maintaining culture viability 9 . However, for enhanced robustness during further processing of this probiotic bacterium, the PEF treatment should be further optimized to increase the fraction of reversible electroporated cells loaded with trehalose 10 .…”

Section: Introductionmentioning

confidence: 99%

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Reversibility of membrane permeabilization upon pulsed electric field treatment in Lactobacillus plantarum WCFS1

Vaessen

Timmermans

Tempelaars

et al. 2019

Sci Rep

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Pulsed electric field (PEF) treatment, or electroporation, can be used to load molecules into cells. The permeabilizing effect of the PEF treatment on the cellular membrane can be either reversible or irreversible depending on the severity of the PEF treatment conditions. The influence of PEF on the reversibility of membrane permeabilization in Lactobacillus plantarum WCFS1 by two different fluorescent staining methods was investigated in this study. Whereas staining with propidium iodide (PI) before and after PEF treatment indicated small reversible permeabilized fractions of maximum 14%, the use of a double staining method with PI and SYTOX Green suggested larger reversible permeabilized fractions up to 40% of the population. This difference shows that the choice for a fluorescent staining method affects the conclusions drawn regarding reversibility of membrane permeabilization. Additionally, the effect of PEF treatment conditions on membrane integrity was compared, indicating a relation between critical electric field strength, cell size and membrane permeabilization. Overall this study showed the possibilities and limitations of fluorescent membrane integrity staining methods for PEF studies.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reversibility of membrane permeabilization upon pulsed electric field treatment in Lactobacillus plantarum WCFS1

Vaessen

Timmermans

Tempelaars

et al. 2019

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Pulse duration usually varies from nanoseconds to a millisecond, and field strength -from several kV/cm to hundreds or thousands of V/cm [14,15]. Apart from the field parameters, the permeability of cellular membranes is affected also by the composition of the buffer used for the electroporation, temperature, and cell properties (size, shape, adherence and density).…”

Section: Introductionmentioning

confidence: 99%

Influence of Pulsed Electric Field on Accumulation of Calcium in Lactobacillus rhamnosus B 442

Góral¹,

Pankiewicz²,

Sujka³

et al. 2020

Journal of Microbiology and Biotechnology

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Calcium is an element that performs many important functions in the human body. A study was conducted on the use of a pulsed electric field (PEF) to enrich cells of Lactobacillus rhamnosus B 442 in calcium ions. The highest concentration of calcium ions in bacterial cells (7.30 mg/g d.m.) was obtained at ion concentration of 200 µg/ml of medium and with the use of the following PEF parameters: field strength 3.0 kV/cm, exposure time 10 min, pulse width 75 ms and 20 h of culturing after which bacteria were treated with the field. Cell biomass varied in the range from 0.09 g/g d.m. to 0.252 g/g d.m., and the total number of bacteria ranged from 10 1 0 CFU/ml to 10 1 2 CFU/ml. Microscope photographs prove that calcium ions were situated within the cells of the bacteria, and electroporation contributed to an increase in the effectiveness of the ion bioaccumulation process. Samples containing calcium and subjected to electroporation displayed intensive fluorescence. The significance of this research was the possibility of using probiotic bacteria enriched with calcium ions for the production of functional food in subsequent studies.

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“…One of the promising technologies to stress microorganisms is PEF. This technology has the ability to induce the formation of pores in the cell membranes and cause the breakdown of the cells, which can be reversible or irreversible, depending mainly on the applied PEF parameters, the medium composition, and the cell type [18,19]. PEF treatments usually involve the application of high intensity pulses (typically > 20 kV cm −1 ) for a treatment time less than 1 s [20].…”

Section: Introductionmentioning

confidence: 99%

Effect of Pulsed Electric Fields on the Growth and Acidification Kinetics of Lactobacillus delbrueckii Subsp. bulgaricus

Peng

Koubàa

Bals

et al. 2020

Foods

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The aim of this work was to investigate the effect of pulsed electric fields (PEF) on the growth and acidification kinetics of Lactobacillus delbrueckii subsp. bulgaricus CFL1 during fermentation. The PEF treatments were applied during the fermentation process using a recirculation pump and a PEF treatment chamber coupled with a PEF generator. The medium flow rate through the chamber was first optimized to obtain the same growth and acidification kinetics than the control fermentation without medium recirculation. Different PEF intensities (60–428 V cm−1) were then applied to the culture medium to study the impact of PEF on the cells’ behavior. The growth and acidification kinetics were recorded during the fermentation and the specific growth rates µ, pH, and acidification rate (dpH/dt) were assessed. The results obtained showed a biphasic growth by applying high PEF intensities (beyond 285 V cm−1) with the presence of two maximal specific growth rates and a decrease in the acidification activities. It was demonstrated that the cells were stressed during the PEF treatment, but presented an accelerated growth after stopping it, leading thereby to similar absorbance and pH at the end of the fermentation. These results show the great potential of PEF technology to be applied to generate low acidified products by performing PEF-assisted fermentations.

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Pulsed electric field for increasing intracellular trehalose content in Lactobacillus plantarum WCFS1

Cited by 28 publications

References 27 publications

Reversibility of membrane permeabilization upon pulsed electric field treatment in Lactobacillus plantarum WCFS1

Reversibility of membrane permeabilization upon pulsed electric field treatment in Lactobacillus plantarum WCFS1

Influence of Pulsed Electric Field on Accumulation of Calcium in Lactobacillus rhamnosus B 442

Effect of Pulsed Electric Fields on the Growth and Acidification Kinetics of Lactobacillus delbrueckii Subsp. bulgaricus

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