The effect of pulsed electric fields on the inactivation and structure of lysozyme

The effect of pulsed electric fields (PEF) on the activity and structure of lysozyme selected as a model enzyme was investigated. The inactivation of lysozyme in phosphate buffer was a function of electric field strength, treatment time, electrical conductivity, and enzyme concentration. No significant (p > 0.05) change in the activity of PEF-treated lysozyme was found after storage for 12, 24 and 48 h at 4 °C. The effect of PEF on tertiary structure of lysozyme was demonstrated by second-derivative UV spectra and intrinsic fluorescence. The results indicated that the unfolding of tertiary structure was induced by PEF treatment at 35 kV/cm for 1200 μs, and more tyrosine residues were buried inside the protein after PEF treatment, accompanied by the exposure of more tryptophan residues. CD spectra suggested that the inactivation of lysozyme by PEF was closely related to the loss of α-helix of secondary structure.     

Assessment of pulsed electric fields induced cellular damage in Saccharomyces cerevisiae: Change in performance of mitochondria and cellular enzymes

Pulsed electric fields (PEF) induced cellular damage in Saccharomyces cerevisiae, mainly focusing on metabolic properties including performance of mitochondria and cellular enzymes, was evaluated in this study. The proportion of sublethally injured S. cerevisiae cells was around 90% after PEF treatment at 20 kV/cm for 100–500 μs. Sublethal injure S. cerevisiae correlated with damage to mitochondria and membranes caused by PEF, which was also confirmed by the glucose-energized repair process. An increase in plasma membrane Ca²⁺-ATPase, decrease of cytoplasmic esterase activity and changes of ten cellular enzymes of S. cerevisiae were observed after PEF treatment, suggesting the sublethal injury of S. cerevisiae is correlated with the inactivation or activation of cellular enzymes and aggregation of proteins under PEF treatment.     

Effects of pulsed electric fields (PEF) treatment on physicochemical properties of potato starch

Potato starch–water suspensions (8.0%, w/w) were subjected to pulsed electric fields (PEF) treatment at 30 kV·cm^− 1, 40 kV·cm^− 1 and 50 kV·cm^− 1, respectively. The physicochemical properties of PEF-treated potato starch samples were investigated using scanning electron microscopy (SEM), laser scattering technique, X-ray diffractometry (XRD), differential scanning calorimetry (DSC), and the Brabender rheological method, with native potato starch as reference. It has been concluded from SEM analysis that dissociation and damage of PEF-treated potato starch granules appeared. Some granules aggregated with each other and showed gel-like structures. It was revealed from particle size analysis that there was an obvious increase of the granule size after PEF treatment. This has been attributed to the aggregation among granules. It was also demonstrated from other analysis that relative crystallinity, gelatinization temperatures, gelatinization enthalpy, peak viscosity as well as breakdown viscosity of modified samples all decreased with increasing electric field strength.

Industrial relevance

In this study, the effect of PEF treatment (up to 50 kV·cm^− 1) on physicochemical properties of potato starch has been investigated. The results from SEM images showed that dissociation, denaturation and damage of potato starch granules had been induced by the PEF treatments. Some of granule fragments showed gel-like structures, and congregated with each other or with other starch granules. Laser scattering measurements of particle size revealed that an obvious increase of granule size under electric field strength of 50 kV·cm^− 1, which was attributed to the aggregation of the starch granules. The X-ray diffraction pattern showed an obvious loss of crystalline structure after the PEF treatment at 50 kV·cm^− 1, which induced a trend of transformation from crystal to non-crystal in potato starch granules. DSC analysis showed a decrease in gelatinization temperatures (T_o and T_p) and gelatinization enthalpy (ΔH_gel) with increasing electric field strength. Brabender rheological method has been used to show that the peak viscosity and breakdown viscosity decrease with increasing electric field strength of PEF treatment. All the results reveal that the PEF treatment can lead to an intragranular molecular rearrangement of potato starch granules, which induces changes of various physicochemical properties of the treated starch thus may endow it some new characteristics and functions. This phenomena may warrant further more detailed study.     

Freezing of potato tissue pre-treated by pulsed electric fields

The effects of pulsed electric fields (PEF) pre-treatment on the freezing, freeze-drying and rehydration behavior of potato were studied. Potato samples (26 mm diameter, 10 mm high) were treated by PEF (400 V/cm) for various durations between 10⁻⁴ and 0.3 s. The degree of tissue damage was quantified by the change in electrical conductivity. PEF treated and untreated samples were either frozen in an air-blast freezer with air at −35 °C and 2 m/s velocity or freeze-dried at 0 °C and 0.04 mbar pressure and then rehydrated in water at 25 °C. The freezing times for PEF pre-treated samples reduced as the PEF-induced tissue damage increased. Scanning electron microscope images of the air-blast frozen and then freeze-dried samples showed increased deformation of cells and larger intercellular spaces (frozen samples only) for the PEF pre-treated samples. However, PEF pre-treatment improved the rate of freeze-drying and improved the quality and rehydration of the samples.     

拉曼光谱研究脉冲电场作用下磷脂脂质体构象的变化

为深入研究脉冲电场的灭菌机理,探讨电场与细胞膜的作用机制,利用拉曼光谱谱图分析了不同电场场强下磷脂脂质体构象的变化。结果表明,拉曼峰值比I1085/I1064随着电场场强的增加而相应增大,而I1085/I1123随着电场场强的增加而先增加后减少,但比值仍高于空白样,说明经过电场处理后,脂质体C-C扭曲构象的数目增多;而I2885/I2850随着场强的增加呈现递减的趋势。由此可以说明经电场处理后的脂质体磷脂分子间烃链的无序度增加,导致细胞膜流动性增强。     

高压脉冲电场对食品中生物大分子的影响

高压脉冲电场非热处理技术因处理时间短、温升小、能耗低和杀菌效果明显等成为目前杀菌工艺中研究最为活跃的技术之一,而且在功能物质提取与保持物质活性等方面也展现了较好的应用前景。着重就高压脉冲电场对食品中的一些生物大分子如酶类、蛋白质等的影响进行综述。     

Emergence of pulsed electric fields resistance in Salmonella enterica serovar Typhimurium SL1344

In this investigation we selected and isolated a culture derived from Salmonella enterica serovar Typhimurium SL1344 with stable increased resistance to pulsed electric fields (PEF) after repeated rounds of PEF treatment and outgrowth of survivors. The resulting culture showed a higher resistance to PEF treatments under different treatment conditions. The acquisition of PEF resistance was only observed in stationary phase cells. The cytoplasmic membrane of the resistant variant showed a higher resilience against PEF treatments, since a lower permeabilization degree was observed after PEF treatments, in comparison to the parental strain. Resistance to PEF was also accompanied by a higher tolerance to acidic pH, hydrogen peroxide and ethanol, but not to heat. The occurrence of a PEF resistant variant in S. enterica serovar Typhimurium SL1344 emphasizes the need to further study the mechanisms of inactivation and resistance by PEF for an adequate design of safe treatments.     

Degradation behaviour of methamidophos and chlorpyrifos in apple juice treated with pulsed electric fields

Apple juice (13 °Brix) spiked with methamidophos and chlorpyrifos (2–3 mg/l of each compound) was treated by pulsed electric fields (PEF), and pesticide residues were quantified by gas chromatography with flame photometric detection (GC-FPD). Results showed that electric field strength (8–20 kV/cm) and pulse number (6–26 pulses) have significant effects on the degradation of methamidophos and chlorpyrifos. PEF treatment is effective for the degradation of methamidophos and chlorpyrifos residues in apple juice, and chlorpyrifos is much more labile to PEF than methamidophos. An increase in either pulse number or electric field strength could speed the degradation of methamidophos and chlorpyrifos, and the kinetics equations and related parameters quantitatively characterized the degradation behavior of the pesticides. The exponential model better fits the experimental data for all treatments than the linear model.     

Reduction of acrylamide formation in potato slices during frying

Reduction of acrylamide formation in potato chips was investigated in relation to frying temperature and three treatments before frying. Potato slices (Tivoli variety, diameter: 37 mm, width: 2.2 mm) were fried at 150°C, 170°C and 190°C until reaching moisture contents of ∼1.7 g water/100 g (total basis). Prior to frying, potato slices were treated in one of the following ways: (i) soaked in distilled water for 0 min (control), 40 min and 90 min; (ii) blanched in hot water at six different time-temperature combinations (50°C for 30 and 70 min; 70°C for 8 and 40 min; 90°C for 2 and 9 min); (iii) immersed in citric acid solutions of different concentrations (10 and 20 g/l) for half an hour. Glucose and asparagine concentration was determined in potato slices before frying, whereas acrylamide content was determined in the resultant fried potato chips. Glucose content decreased in ∼32% in potato slices soaked 90 min in distilled water. Soaked slices showed on average a reduction of acrylamide formation of 27%, 38% and 20% at 150°C, 170°C and 190°C, respectively, when they were compared against the control. Blanching reduced on average 76% and 68% of the glucose and asparagine content compared to the control. Potato slices blanched at 50°C for 70 min surprisingly had a very low acrylamide content (28 μm/kg) even when they were fried at 190°C. Potato immersion in citric acid solutions of 10 and 20 g/l reduced acrylamide formation by almost 70% for slices fried at 150°C. For the three pre-treatments studied, acrylamide formation increased dramatically as the frying temperature increased from 150°C to 190°C.     

The first ESR observation of radical species generated under pulsed electric fields processing

Electron spin resonance (ESR) spectroscopy was used to investigate the generation of free radicals under pulsed electric field (PEF) processing. Spin trapping studies, using 5,5-dimethyl-1-pyrroline N-oxide (DMPO), showed the generation of hydrogen radicals in phosphate buffer and oleic acid emulsion. The concentration of hydrogen peroxide in phosphate buffer after PEF treatment was determined by spectrofluorometric method. Electric field strength has a significant effect on the concentration of H₂O₂ (p < 0.05). In PEF-treated samples with electric field strength of 30 kV/cm and 35 kV/cm, trace concentrations of H₂O₂ (0.177 × 10⁻⁶ and 2.035 × 10⁻⁶ mol/L, respectively) were detected.     

Effect of pulsed electric fields and high pressure on improved recovery of high-added-value compounds from olive pomace

Olive pomace is considered a solid by-product and a rich source of valuable compounds such as polyphenols, flavonoids with antioxidant properties, and proteins. Nonthermal technologies, which cause alterations to cell permeability, are being explored to assist conventional recovery techniques. The aim of this study was to assess the effect of pulsed electric fields (PEF) and high pressure (HP) on improved recovery yield of the high-added-value compounds or to shorten the extraction time of these compounds. Olive pomace (Tsounati cv) was pretreated with PEF (1.0 to 6.5 kV/cm, 0.9 to 51.1 kJ/kg, and 15 µs pulse width) or HP (200 to 600 MPa and 0 to 40 min). Evaluation of the intracellular compounds extracted via solid–liquid extraction (50% ethanol–water solution) was performed. More intense PEF and HP conditions resulted in a significant increase of the phenolic concentration up to 91.6% and 71.8%, respectively. The increased antioxidant capacity of each extract was correlated to phenolic compound concentration. The protein concentration that was achieved with PEF pretreatment was doubled; however, HP-pretreated extracts reached 88.1% higher yield than untreated for pressures up to 200 MPa. HP and PEF pretreatment decreased extraction completion time t₉₈ (needed time to recover the equal amount of phenolics and proteins of untreated after 60 min of conventional extraction) to 12 min and lower than 1 min, respectively. To conclude, both pretreatments are effective in improving the conventional extraction process for increased yield recovery of high-added-value compounds from olive pomace.     

The impact of pulsed electric fields on quality parameters of freeze-dried red beets and pineapples

The effect of pulsed electric fields (PEF) on the cell disintegration index (Zp), the freeze-drying process and the final product quality of red beet and pineapple tissue was studied. Red beets and pineapples were PEF-treated at an electric field strength of 1.07 kV/cm and specific energy inputs of 1 kJ/kg and 4 kJ/kg (PEF₁ and PEF₂, respectively). Freeze-drying was performed at a pressure of 1 mbar and heating plate temperature of 50 °C for red beet and 40 °C for pineapple. The quality of freeze-dried tissue was evaluated by the analysis of residual moisture content, macrostructure, texture, colour and rehydration properties. For freeze-dried red beets, the betalain content from extracts was studied. It could be stated that PEF pre-treatment had no significant influence on drying time reduction, however significantly improved the final quality of freeze-dried product. Moreover, the effect of PEF was strongly depending on type of plant material used.     

脉冲电场和热处理对鲜榨苹果汁贮藏期品质的影响

研究高压脉冲电场处理 (PEF)和热处理对鲜榨苹果汁贮藏期的品质影响。研究发现 ,4℃贮藏 3 0d ,PEF( 2 5kV/cm ,1 0 3 5个脉冲 )和热处理 ( 72℃、1 5s和 90℃、60s)可以降低果汁中微生物数量 ,而且对苹果汁的电导率、Brix和 pH值的影响不显著 (P >0 0 5 )。但是热处理对果汁的浊度有显著影响 (P <0 0 5 ) ,而PEF处理差异不显著 (P >0 0 5 )。PEF处理苹果汁的L值和色值均优于热处理 ,感官评价中风味和色泽比热处理的苹果汁更接近对照的风味和色泽。     

Influence of Pulsed Electric Field and Thermal Treatments on the Quality of Blueberry Juice

Fresh blueberries were selected as trial materials. Two blueberry juice samples were sterilized using pulsed electric field and thermal treatments respectively. Their qualities were analyzed and compared by evaluating the microorganisms in the samples. The changes in the quality of fresh blueberry juice samples during storage were also assessed. The results showed that the inactivation of Escherichia coli increased as electric field strength and treatment time increased. Pulsed electric field treatments at 35 kV/cm and 82 μs reduced E. coli by 5.12 logarithms. Pulsed electric field had less effect on juice quality, but after heat treatment, the vitamin C of blueberry juice sample was reduced by 14.78%, whereas its anthocyanin content was reduced by 3.64%. After sterilization, no significant change was observed in the relative reducing sugar, total acid, phenol contents, and Brix value. The vitamin C and anthocyanins contents of the fresh blueberry juice samples treated with pulsed electric field were higher than those treated with heat sterilization.     

Optimization of betulin extraction process from Inonotus Obliquus with pulsed electric fields

In this work, the extraction of betulin from Inonotus Obliquus applying high intensity pulsed electric fields (PEF) was studied. To extract non-polar compound, 75% ethanol solution was used. The effect of PEF on betulin yield was discussed compared with conventional extracting method. A set of optimal parameters (PEF strength = 40 kV/cm, pulse number = 2, ethanol concentration = 75%, and solid-to-solvent ratio = 1:25) were tested and analyzed.

Industrial relevance

Results of this study show that pulsed electric fields can be used to extract non-polar compound from plant origin raw material, such as Chinese herbal medicine, with easy-to-use equipment, materials and shorter extracting time compared with conventional methods.     

Influence of Pulsed Electric Field on Escherichia coli and Saccharomyces cerevisiae

Heat sterilization destroys the color, smell, taste, and nutrients of food. Pulsed electric field can effectively kill microorganisms for many foods at room temperature without compromising sensory and nutritional quality. The effect of pulsed electric field on the sterilization rates of Escherichia coli and Saccharomyces cerevisiae was investigated. The fundamental mechanism of pulsed electric field sterilization on S. cerevisiae was briefly discussed. The inactivation of E. coli and S. cerevisiae increased as the electric field strength and treatment time increased. Pulsed electric field treatments at 35 kV/cm and 90 μs reduced S. cerevisiae and E. coli by 5.30 and 5.15 log numbers, respectively. The inactivation of E. coli and S. cerevisiae was not remarkable at increased conductivity and the same electric field strength. Concaves and holes in S. cerevisiae cell surface, protoplast deletion, overflow of substances from cells, and denatured DNA were observed. These results supported the hypothesis of “membrane perforation” to explain the mechanism underlying pulsed electric field sterilization.     

脉冲电场对酪蛋白功能性质影响的回复性研究

研究脉冲电场对牛奶酪蛋白功能性质影响的回复性。结果表明：酪蛋白样品经过脉冲电场处理,静置24h 后测定的起泡性能和乳化性能在各处理参数下增加的幅度比经脉冲电场处理后立刻测定的有所降低,并且降低的趋势随着脉冲参数的增大而有所减小。     

Acrylamide in potato crisp—the effect of raw material and processing

Reducing sugars and free amino acids were analysed in slices from three potato cultivars before and after blanching (0-3 min). The potato crisps were deep fried at 185 °C for different times (3-8.5 min), and analysed for the concentration of acrylamide (AA) and moisture. Potato cultivar and the temperature during processing were important parameters for AA formation in potato crisps. The amount increased with an increase in the processing time. Blanching before deep-frying reduced the concentration of free asparagine and reducing sugar in the raw material. We found no effect of blanching as pretreatment on the concentration of AA in the potato crisps. Any relationship was not detected between the levels of asparagine in the different cultivars, before and after blanching, and the formation of AA in the crisp products. However, it was shown that the content of reducing sugars determined the level of AA after frying.     

Influence of SO2 on the consumption of nitrogen compounds through alcoholic fermentation of must sterilized by pulsed electric fields

The aim of this work was to study the influence of SO₂ on the use of nitrogenous compounds by yeast during wine alcoholic fermentation. Thus Parellada must was sterilized by a pulsed electric field treatment and inoculated with Saccharomyces cerevisiae Na33 strain. The fermentations were carried out with SO₂ (20 mg/l) and without SO₂. Results showed that yeast better consumed the amino acids in the first half of fermentation in the presence of SO₂. The final concentration of amino acids in the obtained wine was greater when the must was fermented without SO₂ than when the latter compound was present. Therefore, the presence of SO₂ facilitated the consumption of amino acids and, hence, such wine should have more complex flavour and better microbiological stability than that obtained from fermentation without SO₂.     

Numerical simulations of pulsed electric fields for food preservation: A review

The application of pulsed electric fields is a novel technique to preserve foods in a non-thermal way. One key component of this technology is the treatment chamber, in which the food is exposed to a pulsed electric field to induce permeabilization of biological cells, e.g. to inactivate microorganisms. For a high efficiency of the method and a high product quality a detailed knowledge of the electric field strength and temperature distribution in the chamber is necessary. The numerical simulation of the fluid dynamics coupled with the electric and thermal fields inside the treatment chamber can provide such information with high spacial and temporal resolution. An important goal of the simulations is the optimization of the treatment chamber geometry to improve the uniformity of the electric and thermal fields between the electrodes in order to avoid the over or under-processing of foods or dielectric breakdowns. This article reviews numerical investigations performed on the pulsed electric field process and presents numerical results of a treatment chamber optimization and the solution of coupled fluid dynamical, electrical and thermal problems.

Industrial relevance:

Numerical simulations of the pulsed electric field process provide detailed information of the fluid flow, the temperature and the electric field distributions in treatment chambers under various conditions. Such local information inside the electric field is difficult to obtain experimentally. For a further development of the pulsed electric field technology, numerical simulations can be applied to improve the fundamental understanding of the physical phenomena occurring and to optimize the process with respect to the chamber design and operating conditions.