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Review

Effects of Pulsed Electric Fields and Ultrasound Processing on Proteins and Enzymes: A Review

by
Sai Kranthi Vanga
1,*,
Jin Wang
1,2,*,
Shesha Jayaram
3 and
Vijaya Raghavan
1
1
Department of Bioresource Engineering, Faculty of Agriculture and Environmental Studies, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, Canada
2
Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, China
3
Department of Electrical & Computer Engineering, Faculty of Engineering, University of Waterloo, Waterloo, ON N2L 3G1, Canada
*
Authors to whom correspondence should be addressed.
Processes 2021, 9(4), 722; https://doi.org/10.3390/pr9040722
Submission received: 30 March 2021 / Revised: 15 April 2021 / Accepted: 16 April 2021 / Published: 20 April 2021
(This article belongs to the Special Issue Feature Review Papers in Section "Food Processes")
Versions Notes

Abstract

:
There is increasing demand among consumers for food products free of chemical preservatives, minimally processed and have fresh-like natural flavors. To meet these growing demands, the industries and researchers are finding alternative processing methods, which involve nonthermal methods to obtain a quality product that meets the consumer demands and adheres to the food safety protocols. In the past two decades’ various research groups have developed a wide range of nonthermal processing methods, of which few have shown potential in replacing the traditional thermal processing systems. Among all the methods, ultrasonication (US) and pulsed electric field (PEF) seem to be the most effective in attaining desirable food products. Several researchers have shown that these methods significantly affect various major and minor nutritional components present in food, including proteins and enzymes. In this review, we are going to discuss the effect of nonthermal methods on proteins, including enzymes. This review comprises results from the latest studies conducted from all over the world, which would help the research community and industry investigate the future pathway for nonthermal processing methods, especially in preserving the nutritional safety and integrity of the food.

1. Introduction

Traditional food processing methods generally involve heat transfer through one of the following ways: roasting, drying, boiling, cooking, and frying [1]. These processing methods help extend the shelf life of the product and add variety to the food we consume [1,2]. The earliest recorded remnants of preserving the food through processing dated back to the ancient Egyptian era where “sun drying” was widely prevalent. However, Nicholas Appert was the scientist who successfully demonstrated that food could be preserved for a longer time using thermal techniques. According to the records, he sealed the food in glass jars that were placed in hot water, which destroyed pathogens, thus extending the shelf life of the product [3,4]. With the invention of electricity and industrialization, the scale of operations has increased exponentially to meet the rising population’s demands around the world. Due to the socioeconomic conditions in the post-second-world-war societies, an evident rise in consumption of confectionery, convenience foods and ready-to-eat meals was observed, which are all processed thermally and contain high concentrations of sugars and chemical additives [5,6]. However, the modern-day consumer is asking for more natural and fresh-like foods, which are minimally processed. This rise in demand results from increased awareness of the advantages of consuming fresh foods, especially fruits and vegetables, among the consumers [7]. Reduced risk of cardiovascular diseases and cancer risk are widely associated with the consumption of fresh fruits and vegetables and associated products [8,9,10].
However, not all food products can be consumed raw as they are widely associated with food safety issues, including Salmonella and E Coli O157: H7/O104:H4 outbreaks that can be highly detrimental to health, leading to death in humans [11,12,13]. Further, foods may contain antinutrients that can have adverse effects on health. Antinutrients are the naturally occurring products present in plant foods, which reduce the nutritional availability in the body. They are secondary and tertiary metabolites produced by the plants as part of their natural defense mechanism [14,15,16]. Despite the few advantages of their presence, they are generally undesirable in food products [15]. Both conventional and novel thermal processing methods were found to be highly effective in alleviating both pathogens and antinutrients present in food, and this effectively dealing with the food safety concern [4,14,17]. However, thermal processing influences the sensory and nutritional components in food that can be both advantageous and disadvantageous depending on the various factors, including the food components, processing temperature, duration of processing and method adopted. As mentioned already, the advantages of processing foods thermally include the destruction of pathogens and antinutrients present in food. In few cases, the food can also develop desirable sensory qualities making it more palatable for human consumption [1,6].
Conversely, various studies have also shown the negative impact of thermal treatments in food [18,19]. They were found to degrade various sensitive nutritional components, which include anthocyanins, ascorbic acid and various other bioactive compounds that play a critical role in the wellbeing of the human body [18,19]. They can also lead to undesirable chemical reactions (Ex: Milliard reaction), resulting in undesirable organoleptic and nutritional changes [20,21]. Thus, these downsides of thermal processing have forced the researchers and the food industry to look for alternative processing methods that would preserve the “fresh” like the sensory product, but at the same time provides a desirable effect by eliminating the health risks associated with consumption of raw foods, which lead to developing nonthermal processing methods. Nonthermal processing involves the methods where the food is processed under sublethal temperatures, i.e., the food is not heated up. The rising demand from industries to introduce these methods has put much pressure on researchers to evaluate their effect on various components in food, including proteins [1]. These nonthermal methods were found to give a targeted response based on the application and further can improve the overall sensory and nutritional quality of the food products [20].
Though there are various other major nutritional components present in foods, proteins are the most sensitive and vital for human consumption. They play a significant role in muscle building and maintenance of cell wall structure in humans and plants, respectively. Proteins also act as one of the primary sources of energy (4 kcal/g of protein) [1,22]. Considering the above factors, a considerable amount of research has been done on the effect of thermal processing on proteins which is covered in various reviews published [23,24,25,26,27,28,29]. However, over the past two decades’ considerable amount of progress has been made in assessing the effectiveness of a wide range of nonthermal processing methods in replacing the conventional thermal methods. In this review, the processing effect of electric field applications and ultrasonication on proteins are discussed in detail. This would help both the researchers in assessing the effectiveness of these methods and their influence on both the structural and functional properties of various proteins.

2. Pulsed Electric Field Processing (PEF)

PEF technology is one of the nonthermal processing techniques that has gained much attention in recent years due to its ability to render food safe for consumption by meeting the food safety standards with short treatment time and minimal heat production. Though there are multiple ways of applying electric fields to food, PEF involves using high voltage in short bursts across the food particles placed in between two electrodes [30]. The short bursts last from several nanoseconds to several microseconds, and the applied electric field strength can vary from 10–80 kV/cm. The continuous application of these pulses possesses the ability to inactive pathogens and enzymes, resulting in food quality deterioration while maintaining the fresh-like organoleptic and nutritional properties that consumers demand [31,32,33]. The ability of PEF application to reduce microbial activity has repeatedly been evaluated and discussed extensively [34,35,36,37,38,39,40].
The application of electrical fields to biological materials like various food products results in charge buildup across the cell membrane of the pathogens, which leads to disruption in the normal functioning of the cell wall. In case the potential across this membrane exceeds a critical value, disruption occurs, resulting in the loss of its functional biological properties as a cell membrane causing “cell death” [34,41,42]. The second alternative theory was proposed by Zimmermann [43], which is called ”dielectric rupture theory,” which suggests the cell membrane acts as a capacitor and application of electric field results in the accumulation of charge on the surfaces of the membrane. This accumulation of charge results in an increase of transmembrane potential (TMP), which will, in turn, reduce the thickness of the cell wall. Ultimately, as the value of transmembrane potential reaches a value of around 1 V, pore formation takes place, resulting in damage to the cell. In case the potential is further increased, irreversible damage can occur to the cell resulting in its inactivation [42,43]. The TMP can be calculated using the following equation:
U(t) = 1.5rE
where U(t) is the TMP in the direction of the applied electric field (V); r is the radius of the cell in μm and E is the electric field strength applied in kV/mm [34,38,42].
Tsong [44] also proposed “electroporation theory”, which is based on the ordered structure of the cell membrane and the dipole nature of the lipids present in the membrane. It states that applying an electric field causes electrical and thermal stress resulting in deformation of the lipid conformation leading to creating new hydrophobic pores. These hydrophobic pores are not very stable and hence result in the formation of stable hydrophilic pores. The local heating of the cell membrane results in a thermal transition from gel to the crystalline structure of the lipid bilayer occurs resulting in loss of semipermeable property. Further, the cell wall also contains protein channels that are sensitive to the high voltage applied. In addition, hence applying an electric field can result in denaturation of these sensitive proteins resulting in loss of functional properties [39,42,44].
Apart from the inactivation of pathogens, PEF can also be used for modification of protein secondary structure and functional properties. However, at low intensities, the stress provided may not be sufficient to induce permanent denaturation in various food products [45]. Table 1, Table 2 and Table 3 summarizes the influence of PEF on various proteins and enzymes.
Table 1 summarizes the recent studies that evaluated the structural changes in various proteins. The data shows that the PEF treatment can directly influence the secondary structures in food proteins. Particularly, they reduce the α-helix content of the proteins, which was observed in the case of soy, canola and egg proteins. The electric fields have also resulted in the loss of β-sheets in numerous proteins, including canola, egg and pepsin. However, when the applied electric field is higher (generally over 35 kV/cm), there is a possibility of an increase in the structured conformations, i.e., α-helices and β-sheets, which was observed in soy protein [48]. Table 2 summarizes the changes in physicochemical and functional properties of a wide variety of proteins due to PEF processing. Protein aggregation was found to be common when treated using high-intensity electric fields and for longer processing times. However, pH also seems to be a major factor that influences the protein aggregate formation during PEF processing, as observed in the case of egg white. This must be further evaluated to verify if it can influence aggregation in other proteins, especially plant-based proteins. Furthermore, PEF was found to have no influence on the immunoreactivity of egg and whey proteins (milk) except at very high intensities, which resulted in a slight reduction in the whey protein allergenicity. However, the clinical relevance of this must be further evaluated. The influence of PEF was also evaluated specifically on peanut (Ara h 2,6) and apple (Mal d 3) allergens and showed no significant changes in the secondary structures and reactivity [79]. The impact of PEF on enzyme activity has been outlined in Table 3. The PEF technology was tested on a wide range of beverage products as an alternative to traditional thermal pasteurization. The results presented show that PEF is effective in reducing a wide range of enzymes, including lipoxygenase (LOX), pectinesterase (PE) and peroxidase (PO), in numerous fruit juices and other beverage products.

3. Ultrasonication/Ultrasound Processing (US)

Ultrasound waves are waves with a frequency of 18 kHz and above extending into GHz. They have been employed in the food industry for various applications, such as homogenization, crystallization and emulsification. They are also widely used for the filtration and tenderization of meat [80]. Ultrasound applications of low-energy, i.e., with sound intensities less than 1 W/cm2 and frequencies higher than 100 kHz, are used for non-invasive purposes. They are used as analytical tools for the characterization of physical and chemical properties. Diagnostic ultrasound (1–10 MHz) is one example that uses non-invasive ultrasound waves [80,81]. In the food industry, they are used in surface cleaning, simulation of living cell activity during fermentation through degassing and aid in assessing the physicochemical properties, including particle size, composition and flow rate [81,82]. High-energy ultrasound with sound intensities less than 1 W/cm2 and frequencies in the range of 18–100 kHz are used in food processing areas, including inactivation of enzymes and microbes, degassing, drying, thawing, etc., [82]. Further, applying combinations of heat, pressure and ultrasound have been found to be highly effective, especially in liquid foods. The various forms of ultrasonic applications in food are as follows:

3.1. Presonication

This is the food processing technique where ultrasound is applied to the food product before the actual processing, either by heat or temperature. This method is found to be very effective in reducing the resistance of heat and pressure-resistant microbes and enzymes [83].

3.2. Postsonication

In postsonication, the product that is treated either by heat or pressure is then subjected to ultrasound. There is no experimental data on this method as the method is not widely used in the process industry [84].

3.3. Thermosonication

In this method, both ultrasonic waves and heat (moderate levels) are applied simultaneously. The process of thermosonication is found to be particularly effective in the inactivation of vegetative cells and enzymes [84].

3.4. Manosonication

The processing is performed by subjecting food to both ultrasonic waves and medium pressures in the range of 100–1000 kPa at low temperatures [83,84].

3.5. Manothermosonication

Manothermosonication (MTS) is applying a combination of temperature, pressure and ultrasound on a product simultaneously. Temperatures can range between (30–140 °C), and the pressures can go up to 1000 kPa in this processing method, which was found to be very effective for treating yeasts and various enzymes in liquid foods [83,84].
The first use of ultrasonic waves was for the inactivation of microorganisms and was published in the late 1920s [85,86]. Through continuous work has been done on its effectiveness on various pathogens in food, its use was not as widespread as thermal due to the limitations in terms of equipment. But, there was a revival in its application in the food industry in the two decades due to its efficacy in dealing with both unwanted enzymes and microbes in various food products without affecting the nutritional properties as in the case of thermal processing methods. It was also found to have various secondary effects in food, as already mentioned, and since it has been adopted in various process industries [86]. There is a considerable amount of work done by researchers to evaluate the efficacy of ultrasonic processing, especially in combination with thermal processing and nonthermal processing methods like elevated pressures and pH variations. The inactivation effect was caused due to the cavitation, i.e., inception, evolution and implosion of small gas bubbles within the target food matrix. The implosion occurs rapidly and violently, leading to extreme temperatures and pressures on a microscale [87,88]. The external stress is caused due to cavitation was found to be effective in the breakdown of hydrogen bonds leading to conformational changes in tertiary and secondary structure [89]. Further, the cavitation also caused free radical release due to the hemolytic cleavage of water. The hydrogen and hydroxyl free radicals may react with free amino acids that result in the destabilization of enzymes and proteins, leading to changes in the biological activity of the compounds [88,90].
Ultrasonication has been considered as one of the green food processing technologies, which has caught the attention of scientists because of its advantages in effective mass transfer and energy usage, low cost and process temperatures [91,92,93,94]. The mechanism of ultrasonication is due to the formation of cavitation effects during food processing. Cavitation is a phenomenon where the movement of high-power sound waves results in the inception of members of gas/vapor bubbles that progress and then implode within the sample/solvent. This implosion results in extreme conditions of high temperatures (upwards of 1000 °C) and pressures (50–500 MPa) [91,92,95,96]. According to the frequency ranges, low-intensity ultrasonication (0–1 W/cm2, >100 kHz) and high-intensity ultrasonication (>1 W/cm2, 100–200 kHz) are two common types of ultrasonication [92]. Among them, low-intensity ultrasonication is used as a nondestructive tool to monitor the changes of physicochemical compounds during food processing. High-intensity ultrasonication can be used to inhibit the activity of enzymes and microorganisms from extending the shelf life of food products and modifying the secondary structure changes in the proteins leading to change in their functional properties and nutritional value [92,93,96].
As shown in Table 4, the impact of ultrasonication on the structural properties of a wide range of proteins has been outlined. Many studies have reported that the changes in the secondary structures observed due to ultrasonication were found to be highly dependent on the protein itself. The α-helix content in various milk proteins was found to increase after high-intensity sonication, whereas the α-helices reduced in the case of black bean protein and soybean protein [91,97,98,99,100]. In the case of β-sheets content, it reduced in case of protein from milk and soy, while it increased in black bean protein. Protein aggregation was also observed in various proteins that were ultrasonicated [93,101]. The changes in the functional properties of proteins are shown in Table 5. Protein solubility increased in most of the food products, including milk, soymilk, black-bean protein and peanut proteins, with the exception of egg proteins [97,98,102]. In addition, a study found that 16 min US treatment reduced trypsin inhibitor activity of soymilk by 52% [95]. Foamability and emulsification properties have also been enhanced in various plant-based proteins, including wheat, faba bean and plum seed protein [100,103,104]. Ultrasonication was also found to reduce the allergenicity of shrimp, peanut and kiwifruit proteins significantly when compared to untreated control [94,96,105,106]. As shown in Table 6, the effect of US processing on enzymes from a wide range of sources was described. Studies found that US processing can effectively inhibit the activity of polyphenol oxidase (PPO), peroxidase (POD), and pectin methylesterase (PME) present in tomato juice, apple juice, cantaloupe melon juice, pear juice and milk [107,108,109,110,111]. It can be concluded that this nonthermal processing technique possesses the ability to effectively reduce the enzymatic activity that can promote the shelf stability of these beverage products.

4. Conclusions

The primary objective was to evaluate the impact of PEF and US processing on proteins and enzymes of various food products. There has been a growing need to address the consumer demand for “minimally processed” foods, which can be achieved by nonthermal processing techniques like PEF and US processing. Though their fundamental mode of action on food is different, both the methods evaluated were found to be effective in inactivating various enzymes in fruit juices and improve their shelf life. There were changes observed in the secondary and tertiary structures of proteins in many food products. However, these changes were highly dependent on the intensity of processing, the local-food matrix and properties of the protein present in the given product. In some cases, these modifications in their conformational structures have resulted in physicochemical and functional property changes. High-intensity PEF was found to cause protein aggregation in most cases, while ultrasonication was found to increase proteins’ solubility despite aggregation when processed for longer durations. Both PEF and US processing methods show great potential in improving the allergenicity of certain food products. However, further, reach is warranted to understand the clinical implications. Moreover, it is remarkable that researchers are exploring using the combination of these technologies and their impact on various nutritional components [128,129]. Further emphasis should be placed on understanding the combination treatment on proteins and enzymes.

Author Contributions

Conceptualization, S.K.V. and J.W.; methodology, S.K.V.; data curation, S.K.V. and J.W.; writing—original draft preparation, S.K.V. and J.W; writing—review and editing, S.J. and V.R.; supervision, S.J. and V.R; project administration, V.R.; funding acquisition, V.R. All authors have read and agreed to the published version of the manuscript.

Funding

The authors are grateful to the discovery grant provided by NSERC (Natural Sciences and Engineering Research Council of Canada) as financial support for the research program executed by the Department of Bioresource Engineering.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Vanga, S.K.; Singh, A.; Raghavan, V. Review of conventional and novel food processing methods on food allergens. Crit. Rev. Food Sci. Nutr. 2017, 57, 2077–2094. [Google Scholar] [CrossRef]
  2. Dewanto, V.; Wu, X.; Adom, K.K.; Liu, R.H. Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. J. Agric. Food Chem. 2002, 50, 3010–3014. [Google Scholar] [CrossRef] [PubMed]
  3. Vanga, S.K.; Raghavan, V. Processing effects on tree nut allergens: A review. Crit. Rev. Food Sci. Nutr. 2017, 57, 3794–3806. [Google Scholar] [CrossRef] [PubMed]
  4. Stumbo, C.R. Thermobacteriology in Food Processing; Elsevier: New York, NY, USA, 2013. [Google Scholar]
  5. Van Boekel, M.; Fogliano, V.; Pellegrini, N.; Stanton, C.; Scholz, G.; Lalljie, S.; Somoza, V.; Knorr, D.; Jasti, P.R.; Eisenbrand, G. A review on the beneficial aspects of food processing. Mol. Nutr. Food Res. 2010, 54, 1215–1247. [Google Scholar] [CrossRef]
  6. Fellows, P.J. Food Processing Technology: Principles and Practice; Elsevier: Cambridge, UK, 2009. [Google Scholar]
  7. Kaur, C.; Kapoor, H.C. Antioxidants in fruits and vegetables—The millennium’s health. Int. J. Food Sci. Technol. 2001, 36, 703–725. [Google Scholar] [CrossRef]
  8. Ness, A.R.; Powles, J.W. Fruit and vegetables, and cardiovascular disease: A review. Int. J. Epidemiol. 1997, 26, 1–13. [Google Scholar] [CrossRef] [Green Version]
  9. Block, G.; Patterson, B.; Subar, A. Fruit, vegetables, and cancer prevention: A review of the epidemiological evidence. Nutr. Cancer 1992, 18, 1–29. [Google Scholar] [CrossRef]
  10. Steinmetz, K.A.; Potter, J.D. Vegetables, fruit, and cancer prevention: A review. J. Am. Diet. Assoc. 1996, 96, 1027–1039. [Google Scholar] [CrossRef]
  11. Karch, H.; Denamur, E.; Dobrindt, U.; Finlay, B.B.; Hengge, R.; Johannes, L.; Ron, E.Z.; Tønjum, T.; Sansonetti, P.J.; Vicente, M. The enemy within us: Lessons from the 2011 european escherichia coli o104:H4 outbreak. EMBO Mol. Med. 2012, 4, 841–848. [Google Scholar] [CrossRef]
  12. Besser, R.E.; Lett, S.M.; Weber, J.T.; Doyle, M.P.; Barrett, T.J.; Wells, J.G.; Griffin, P.M. An outbreak of diarrhea and hemolytic uremic syndrome from escherichia coli o157: H7 in fresh-pressed apple cider. JAMA 1993, 269, 2217–2220. [Google Scholar] [CrossRef] [PubMed]
  13. Beuchat, L.R. Pathogenic microorganisms associated with fresh produce. J. Food Prot. 1996, 59, 204–216. [Google Scholar] [CrossRef]
  14. Vagadia, B.H.; Vanga, S.K.; Singh, A.; Raghavan, V. Effects of thermal and electric fields on soybean trypsin inhibitor protein: A molecular modelling study. Innov. Food Sci. Emerg. Technol. 2016, 35, 9–20. [Google Scholar] [CrossRef]
  15. Thompson, L.U. Potential health benefits and problems associated with antinutrients in foods. Food Res. Int. 1993, 26, 131–149. [Google Scholar] [CrossRef]
  16. Akande, K.; Doma, U.; Agu, H.; Adamu, H. Major antinutrients found in plant protein sources: Their effect on nutrition. Pak. J. Nutr. 2010, 9, 827–832. [Google Scholar] [CrossRef] [Green Version]
  17. Lewis, M.; Heppell, N.; Hastings, A. Continuous Thermal Processing of Foods—Pasteurization and uht Sterilization; Aspen Publishers: New York, NY, USA, 2001. [Google Scholar]
  18. Patras, A.; Brunton, N.P.; O’Donnell, C.; Tiwari, B. Effect of thermal processing on anthocyanin stability in foods; mechanisms and kinetics of degradation. Trends Food Sci. Technol. 2010, 21, 3–11. [Google Scholar] [CrossRef]
  19. Lathrop, P.; Leung, H. Rates of ascorbic acid degradation during thermal processing of canned peas. J. Food Sci. 1980, 45, 152–153. [Google Scholar] [CrossRef]
  20. Knorr, D.; Ade-Omowaye, B.I.O.; Heinz, V. Nutritional improvement of plant foods by non-thermal processing. Proc. Nutr. Soc. 2002, 61, 311–318. [Google Scholar] [CrossRef] [Green Version]
  21. Ohlsson, T. Minimal processing-preservation methods of the future: An overview. Trends Food Sci. Technol. 1994, 5, 341–344. [Google Scholar] [CrossRef]
  22. Phillips, G.O.; Williams, P.A. Handbook of Food Proteins; Woodhead Publishing: Cambridge, UK, 2011. [Google Scholar]
  23. Davis, P.; Williams, S. Protein modification by thermal processing. Allergy 1998, 53, 102–105. [Google Scholar] [CrossRef]
  24. Davis, P.J.; Smales, C.M.; James, D.C. How can thermal processing modify the antigenicity of proteins? Allergy 2001, 56, 56–60. [Google Scholar] [CrossRef]
  25. Wijayanti, H.B.; Bansal, N.; Deeth, H.C. Stability of whey proteins during thermal processing: A review. Compr. Rev. Food Sci. Food Saf. 2014, 13, 1235–1251. [Google Scholar] [CrossRef]
  26. Back, J.F.; Oakenfull, D.; Smith, M.B. Increased thermal stability of proteins in the presence of sugars and polyols. Biochemistry 1979, 18, 5191–5196. [Google Scholar] [CrossRef]
  27. Bahar, I.; Atilgan, A.R.; Erman, B. Direct evaluation of thermal fluctuations in proteins using a single-parameter harmonic potential. Fold. Des. 1997, 2, 173–181. [Google Scholar] [CrossRef] [Green Version]
  28. Ahnoff, M.; Cazares, L.H.; Sköld, K. Thermal inactivation of enzymes and pathogens in biosamples for ms analysis. Bioanalysis 2015, 7, 1885–1899. [Google Scholar] [CrossRef]
  29. Mondoulet, L.; Paty, E.; Drumare, M.F.; Ah-Leung, S.; Scheinmann, P.; Willemot, R.M.; Wal, J.M.; Bernard, H. Influence of thermal processing on the allergenicity of peanut proteins. J. Agric. Food Chem. 2005, 53, 4547–4553. [Google Scholar] [CrossRef]
  30. Barbosa-Cánovas, G.; Gongora-Nieto, M.; Pothakamury, U.; Swanson, B. Fundamentals of high-intensity pulsed electric fields (pef). In Preservation of Foods with Pulsed Electric Fields; Acadamic Press: San Diego, CA, USA, 1999; pp. 1–19. [Google Scholar]
  31. Barba, F.J.; Parniakov, O.; Pereira, S.A.; Wiktor, A.; Grimi, N.; Boussetta, N.; Saraiva, J.A.; Raso, J.; Martin-Belloso, O.; Witrowa-Rajchert, D.; et al. Current applications and new opportunities for the use of pulsed electric fields in food science and industry. Food Res. Int. 2015, 77, 773–798. [Google Scholar] [CrossRef]
  32. Jayaram, S.H. Sterilization of liquid foods by pulsed electric fields. IEEE Electr. Insul. Mag. 2000, 16, 17–25. [Google Scholar] [CrossRef]
  33. Walkling-Ribeiro, M.; Rodríguez-González, O.; Jayaram, S.; Griffiths, M.W. Microbial inactivation and shelf life comparison of ‘cold’ hurdle processing with pulsed electric fields and microfiltration, and conventional thermal pasteurisation in skim milk. Int. J. Food Microbiol. 2011, 144, 379–386. [Google Scholar] [CrossRef] [PubMed]
  34. Sale, A.; Hamilton, W. Effects of high electric fields on microorganisms: I. Killing of bacteria and yeasts. Biochim. Biophys. Acta (BBA)-Gen. Subj. 1967, 148, 781–788. [Google Scholar] [CrossRef]
  35. Wang, M.-S.; Wang, L.-H.; Bekhit, A.E.-D.A.; Yang, J.; Hou, Z.-P.; Wang, Y.-Z.; Dai, Q.-Z.; Zeng, X.-A. A review of sublethal effects of pulsed electric field on cells in food processing. J. Food Eng. 2018, 223, 32–41. [Google Scholar] [CrossRef]
  36. Huang, K.; Tian, H.; Gai, L.; Wang, J. A review of kinetic models for inactivating microorganisms and enzymes by pulsed electric field processing. J. Food Eng. 2012, 111, 191–207. [Google Scholar] [CrossRef]
  37. Zhang, Q.; Barbosa-Cánovas, G.V.; Swanson, B.G. Engineering aspects of pulsed electric field pasteurization. J. Food Eng. 1995, 25, 261–281. [Google Scholar] [CrossRef]
  38. Hamilton, W.; Sale, A. Effects of high electric fields on microorganisms: Ii. Mechanism of action of the lethal effect. Biochim. Biophys. Acta (BBA)-Gen. Subj. 1967, 148, 789–800. [Google Scholar] [CrossRef]
  39. Wouters, P.C.; Alvarez, I.; Raso, J. Critical factors determining inactivation kinetics by pulsed electric field food processing. Trends Food Sci. Technol. 2001, 12, 112–121. [Google Scholar] [CrossRef]
  40. Wouters, P.C.; Dutreux, N.; Smelt, J.P.P.M.; Lelieveld, H.L.M. Effects of pulsed electric fields on inactivation kinetics of listeria innocua. Appl. Environ. Microbiol. 1999, 65, 5364–5371. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  41. Martín-Belloso, O.; Soliva-Fortuny, R. Pulsed electric fields processing basics. In Nonthermal Processing Technologies for Food; Wiley-Blackwell: Hoboken, NJ, USA, 2010; pp. 155–175. [Google Scholar]
  42. Jeyamkondan, S.; Jayas, D.S.; Holley, R.A. Pulsed electric field processing of foods: A review. J. Food Prot. 1999, 62, 1088–1096. [Google Scholar] [CrossRef]
  43. Zimmermann, U. Electrical breakdown, electropermeabilization and electrofusion. In Reviews of Physiology, Biochemistry and Pharmacology; Springer: New York, NY, USA, 1986; Volume 105, pp. 175–256. [Google Scholar]
  44. Tsong, T.Y. Electroporation of cell membranes. Biophys. J. 1991, 60, 297–306. [Google Scholar] [CrossRef] [Green Version]
  45. Han, Z.; Cai, M.-J.; Cheng, J.-H.; Sun, D.-W. Effects of electric fields and electromagnetic wave on food protein structure and functionality: A review. Trends Food Sci. Technol. 2018, 75, 1–9. [Google Scholar] [CrossRef]
  46. Zhong, K.; Hu, X.; Zhao, G.; Chen, F.; Liao, X. Inactivation and conformational change of horseradish peroxidase induced by pulsed electric field. Food Chem. 2005, 92, 473–479. [Google Scholar] [CrossRef]
  47. Li, Y.-Q. Structure changes of soybean protein isolates by pulsed electric fields. Phys. Procedia 2012, 33, 132–137. [Google Scholar] [CrossRef] [Green Version]
  48. Liu, Y.Y.; Zeng, X.A.; Deng, Z.; Yu, S.J.; Yamasaki, S. Effect of pulsed electric field on the secondary structure and thermal properties of soy protein isolate. Eur. Food Res. Technol. 2011, 233, 841. [Google Scholar] [CrossRef]
  49. Zhao, W.; Yang, R. The effect of pulsed electric fields on the inactivation and structure of lysozyme. Food Chem. 2008, 110, 334–343. [Google Scholar] [CrossRef]
  50. Li, Y.; Chen, Z.; Mo, H. Effects of pulsed electric fields on physicochemical properties of soybean protein isolates. LWT-Food Sci. Technol. 2007, 40, 1167–1175. [Google Scholar] [CrossRef]
  51. Perez, O.E.; Pilosof, A.M.R. Pulsed electric fields effects on the molecular structure and gelation of β-lactoglobulin concentrate and egg white. Food Res. Int. 2004, 37, 102–110. [Google Scholar] [CrossRef]
  52. Yang, W.; Tu, Z.; Wang, H.; Zhang, L.; Gao, Y.; Li, X.; Tian, M. Immunogenic and structural properties of ovalbumin treated by pulsed electric fields. Int. J. Food Prop. 2017, 20, S3164–S3176. [Google Scholar] [CrossRef]
  53. Qian, J.-Y.; Ma, L.-J.; Wang, L.-J.; Jiang, W. Effect of pulsed electric field on structural properties of protein in solid state. LWT-Food Sci. Technol. 2016, 74, 331–337. [Google Scholar] [CrossRef]
  54. Yang, R.; Li, S.-Q.; Zhang, Q.H. Effects of pulsed electric fields on the activity and structure of pepsin. J. Agric. Food Chem. 2004, 52, 7400–7406. [Google Scholar] [CrossRef] [PubMed]
  55. Zhao, W.; Yang, R. Effect of high-intensity pulsed electric fields on the activity, conformation and self-aggregation of pepsin. Food Chem. 2009, 114, 777–781. [Google Scholar] [CrossRef]
  56. Xiang, B.; Ngadi, M.; Ochoa-Martinez, L.; Simpson, M. Pulsed electric field-induced structural modification of whey protein isolate. Food Bioprocess Technol. 2011, 4, 1341–1348. [Google Scholar] [CrossRef]
  57. Zhang, L.; Wang, L.-J.; Jiang, W.; Qian, J.-Y. Effect of pulsed electric field on functional and structural properties of canola protein by pretreating seeds to elevate oil yield. LWT 2017, 84, 73–81. [Google Scholar] [CrossRef]
  58. Dong, M.; Xu, Y.; Zhang, Y.; Han, M.; Wang, P.; Xu, X.; Zhou, G. Physicochemical and structural properties of myofibrillar proteins isolated from pale, soft, exudative (pse)-like chicken breast meat: Effects of pulsed electric field (pef). Innov. Food Sci. Emerg. Technol. 2020, 59, 102277. [Google Scholar] [CrossRef]
  59. Liu, Y.-F.; Oey, I.; Bremer, P.; Carne, A.; Silcock, P. Effects of ph, temperature and pulsed electric fields on the turbidity and protein aggregation of ovomucin-depleted egg white. Food Res. Int. 2017, 91, 161–170. [Google Scholar] [CrossRef]
  60. Sui, Q.; Roginski, H.; Williams, R.P.; Versteeg, C.; Wan, J. Effect of pulsed electric field and thermal treatment on the physicochemical and functional properties of whey protein isolate. Int. Dairy J. 2011, 21, 206–213. [Google Scholar] [CrossRef]
  61. Luis, R.D.; Arias, O.; Puértolas, E.; Benedé, S.; Sanchez, L.; Calvo, M.; Pérez, M.D. Effect of high-intensity pulse electric fields on denaturation of bovine whey proteins. Milchwissenschaft 2009, 64, 422–426. [Google Scholar]
  62. Mathys, A.; Toepfl, S.; Siemer, C.; Favre, L.; Benyacoub, J.; Hansen, C.E. Pulsed Electric Field Treatment Process and Dairy Product Comprising Bioactive Molecules Obtainable by the Process. U.S. Patent Application 14/131,268, 2 June 2014. [Google Scholar]
  63. Sharma, P.; Oey, I.; Everett, D.W. Effect of pulsed electric field processing on the functional properties of bovine milk. Trends Food Sci. Technol. 2014, 35, 87–101. [Google Scholar] [CrossRef]
  64. Aguiló-Aguayo, I.; Soliva-Fortuny, R.; Martín-Belloso, O. Changes in viscosity and pectolytic enzymes of tomato and strawberry juices processed by high-intensity pulsed electric fields. Int. J. Food Sci. Technol. 2009, 44, 2268–2277. [Google Scholar] [CrossRef]
  65. Manzoor, M.F.; Ahmad, N.; Aadil, R.M.; Rahaman, A.; Ahmed, Z.; Rehman, A.; Siddeeg, A.; Zeng, X.A.; Manzoor, A. Impact of pulsed electric field on rheological, structural, and physicochemical properties of almond milk. J. Food Process Eng. 2019, 42, e13299. [Google Scholar] [CrossRef]
  66. Li, Y.-Q.; Chen, Q.; Liu, X.-H.; Chen, Z.-X. Inactivation of soybean lipoxygenase in soymilk by pulsed electric fields. Food Chem. 2008, 109, 408–414. [Google Scholar] [CrossRef] [PubMed]
  67. Riener, J.; Noci, F.; Cronin, D.A.; Morgan, D.J.; Lyng, J.G. Combined effect of temperature and pulsed electric fields on soya milk lipoxygenase inactivation. Eur. Food Res. Technol. 2008, 227, 1461–1465. [Google Scholar] [CrossRef]
  68. Min, S.; Min, S.K.; Zhang, Q.H. Inactivation kinetics of tomato juice lipoxygenase by pulsed electric fields. J. Food Sci. 2003, 68, 1995–2001. [Google Scholar] [CrossRef]
  69. Aguiló-Aguayo, I.; Soliva-Fortuny, R.; Martín-Belloso, O. Effects of high-intensity pulsed electric fields on lipoxygenase and hydroperoxide lyase activities in tomato juice. J. Food Sci. 2009, 74, C595–C601. [Google Scholar] [CrossRef]
  70. Aguiló-Aguayo, I.; Odriozola-Serrano, I.; Quintão-Teixeira, L.J.; Martín-Belloso, O. Inactivation of tomato juice peroxidase by high-intensity pulsed electric fields as affected by process conditions. Food Chem. 2008, 107, 949–955. [Google Scholar] [CrossRef]
  71. Van Loey, A.; Verachtert, B.; Hendrickx, M. Effects of high electric field pulses on enzymes. Trends Food Sci. Technol. 2001, 12, 94–102. [Google Scholar] [CrossRef]
  72. Aguiló-Aguayo, I.; Soliva-Fortuny, R.; Martín-Belloso, O. Impact of high-intensity pulsed electric field variables affecting peroxidase and lipoxygenase activities of watermelon juice. LWT-Food Sci. Technol. 2010, 43, 897–902. [Google Scholar] [CrossRef]
  73. Riener, J.; Noci, F.; Cronin, D.A.; Morgan, D.J.; Lyng, J.G. Combined effect of temperature and pulsed electric fields on apple juice peroxidase and polyphenoloxidase inactivation. Food Chem. 2008, 109, 402–407. [Google Scholar] [CrossRef]
  74. Marsellés-Fontanet, Á.R.; Martín-Belloso, O. Optimization and validation of pef processing conditions to inactivate oxidative enzymes of grape juice. J. Food Eng. 2007, 83, 452–462. [Google Scholar] [CrossRef]
  75. Shamsi, K.; Versteeg, C.; Sherkat, F.; Wan, J. Alkaline phosphatase and microbial inactivation by pulsed electric field in bovine milk. Innov. Food Sci. Emerg. Technol. 2008, 9, 217–223. [Google Scholar] [CrossRef]
  76. Sharma, P.; Oey, I.; Everett, D.W. Thermal properties of milk fat, xanthine oxidase, caseins and whey proteins in pulsed electric field-treated bovine whole milk. Food Chem. 2016, 207, 34–42. [Google Scholar] [CrossRef]
  77. Elez-Martinez, P.; Suarez-Recio, M.; Martin-Belloso, O. Modeling the reduction of pectin methyl esterase activity in orange juice by high intensity pulsed electric fields. J. Food Eng. 2007, 78, 184–193. [Google Scholar] [CrossRef]
  78. Riener, J.; Noci, F.; Cronin, D.A.; Morgan, D.J.; Lyng, J.G. Combined effect of temperature and pulsed electric fields on pectin methyl esterase inactivation in red grapefruit juice (citrus paradisi). Eur. Food Res. Technol. 2009, 228, 373–379. [Google Scholar] [CrossRef]
  79. Johnson, P.E.; Van Der Plancken, I.; Balasa, A.; Husband, F.A.; Grauwet, T.; Hendrickx, M.; Knorr, D.; Mills, E.N.C.; Mackie, A.R. High pressure, thermal and pulsed electric-field-induced structural changes in selected food allergens. Mol. Nutr. Food Res. 2010, 54, 1701–1710. [Google Scholar] [CrossRef]
  80. Knorr, D.; Zenker, M.; Heinz, V.; Lee, D.-U. Applications and potential of ultrasonics in food processing. Trends Food Sci. Technol. 2004, 15, 261–266. [Google Scholar] [CrossRef]
  81. Patist, A.; Bates, D. Ultrasonic innovations in the food industry: From the laboratory to commercial production. Innov. Food Sci. Emerg. Technol. 2008, 9, 147–154. [Google Scholar] [CrossRef]
  82. Tao, Y.; Sun, D.-W. Enhancement of food processes by ultrasound: A review. Crit. Rev. Food Sci. Nutr. 2015, 55, 570–594. [Google Scholar] [CrossRef]
  83. Kuldiloke, J. Effect of Ultrasound, Temperature and Pressure Treatments on Enzyme Activity and Quality Indicators of Fruit and Vegetable Juices; Technischen Universität: Berlin, Germany, 2002. [Google Scholar]
  84. Kamani, M.H.; Sabahi, S.; Eslami, N.; Azizi, S. Investigating the Usage of Manothermosonication Nonthermal Energy in Food Processing. Int. Res. J. of App. Basic Sci. 2015, 9, 913–916. [Google Scholar]
  85. Harvey, E.N.; Loomis, A.L. The destruction of luminous bacteria by high frequency sound waves. J. Bacteriol. 1929, 17, 373. [Google Scholar] [CrossRef] [Green Version]
  86. Mason, T.J.; Riera, E.; Vercet, A.; Lopez-Buesa, P. 13-application of ultrasound a2-sun, da-wen. In Emerging Technologies for Food Processing; Academic Press: London, UK, 2005; pp. 323–351. [Google Scholar]
  87. Zinoviadou, K.G.; Galanakis, C.M.; Brnčić, M.; Grimi, N.; Boussetta, N.; Mota, M.J.; Saraiva, J.A.; Patras, A.; Tiwari, B.; Barba, F.J. Fruit juice sonication: Implications on food safety and physicochemical and nutritional properties. Food Res. Int. 2015, 77, 743–752. [Google Scholar] [CrossRef]
  88. Shiferaw Terefe, N.; Buckow, R.; Versteeg, C. Quality-related enzymes in plant-based products: Effects of novel food-processing technologies part 3: Ultrasonic processing. Crit. Rev. Food Sci. Nutr. 2015, 55, 147–158. [Google Scholar] [CrossRef] [PubMed]
  89. Lopez, P.; Sala, F.J.; de la Fuente, J.L.; Condon, S.; Raso, J.; Burgos, J. Inactivation of peroxidase, lipoxygenase, and polyphenol oxidase by manothermosonication. J. Agric. Food Chem. 1994, 42, 252–256. [Google Scholar] [CrossRef]
  90. Tian, Z.M.; Wan, M.X.; Wang, S.P.; Kang, J.Q. Effects of ultrasound and additives on the function and structure of trypsin. Ultrason. Sonochem. 2004, 11, 399–404. [Google Scholar] [CrossRef] [PubMed]
  91. Vanga, S.K.; Wang, J.; Orsat, V.; Raghavan, V. Effect of pulsed ultrasound, a green food processing technique, on the secondary structure and in-vitro digestibility of almond milk protein. Food Res. Int. 2020, 137, 109523. [Google Scholar] [CrossRef] [PubMed]
  92. Wang, J.; Wang, J.; Vanga, S.K.; Raghavan, V. High-intensity ultrasound processing of kiwifruit juice: Effects on the microstructure, pectin, carbohydrates and rheological properties. Food Chem. 2020, 313, 126121. [Google Scholar] [CrossRef]
  93. Zhu, Y.; Vanga, S.K.; Wang, J.; Raghavan, V. Effects of ultrasonic and microwave processing on avidin assay and secondary structures of egg white protein. Food Bioprocess Technol. 2018, 11, 1974–1984. [Google Scholar] [CrossRef]
  94. Dong, X.; Wang, J.; Raghavan, V. Effects of high-intensity ultrasound processing on the physiochemical and allergenic properties of shrimp. Innov. Food Sci. Emerg. Technol. 2020, 65, 102441. [Google Scholar] [CrossRef]
  95. Vanga, S.K.; Wang, J.; Raghavan, V. Effect of ultrasound and microwave processing on the structure, in-vitro digestibility and trypsin inhibitor activity of soymilk proteins. LWT 2020, 131, 109708. [Google Scholar] [CrossRef]
  96. Wang, J.; Wang, J.; Vanga, S.K.; Raghavan, V. Influence of high-intensity ultrasound on the ige binding capacity of act d 2 allergen, secondary structure, and in-vitro digestibility of kiwifruit proteins. Ultrason. Sonochem. 2021, 71, 105409. [Google Scholar] [CrossRef]
  97. Villamiel, M.; de Jong, P. Influence of high-intensity ultrasound and heat treatment in continuous flow on fat, proteins, and native enzymes of milk. J. Food Chem. 2000, 48, 472–478. [Google Scholar] [CrossRef]
  98. Yanjun, S.; Jianhang, C.; Shuwen, Z.; Hongjuan, L.; Jing, L.; Lu, L.; Uluko, H.; Yanling, S.; Wenming, C.; Wupeng, G.; et al. Effect of power ultrasound pre-treatment on the physical and functional properties of reconstituted milk protein concentrate. J. Food Eng. 2014, 124, 11–18. [Google Scholar] [CrossRef]
  99. Stanic-Vucinic, D.; Stojadinovic, M.; Atanaskovic-Markovic, M.; Ognjenovic, J.; Grönlund, H.; van Hage, M.; Lantto, R.; Sancho, A.I.; Velickovic, T.C. Structural changes and allergenic properties of β-lactoglobulin upon exposure to high-intensity ultrasound. Mol. Nutr. Food Res. 2012, 56, 1894–1905. [Google Scholar] [CrossRef]
  100. Jiang, L.; Wang, J.; Li, Y.; Wang, Z.; Liang, J.; Wang, R.; Chen, Y.; Ma, W.; Qi, B.; Zhang, M. Effects of ultrasound on the structure and physical properties of black bean protein isolates. Food Res. Int. 2014, 62, 595–601. [Google Scholar] [CrossRef]
  101. Martínez-Velasco, A.; Lobato-Calleros, C.; Hernández-Rodríguez, B.E.; Román-Guerrero, A.; Alvarez-Ramirez, J.; Vernon-Carter, E.J. High intensity ultrasound treatment of faba bean (vicia faba l.) protein: Effect on surface properties, foaming ability and structural changes. Ultrason. Sonochem. 2018, 44, 97–105. [Google Scholar] [CrossRef]
  102. Jambrak, A.R.; Lelas, V.; Mason, T.J.; Krešić, G.; Badanjak, M. Physical properties of ultrasound treated soy proteins. J. Food Eng. 2009, 93, 386–393. [Google Scholar] [CrossRef]
  103. Jambrak, A.R.; Mason, T.J.; Lelas, V.; Herceg, Z.; Herceg, I.L. Effect of ultrasound treatment on solubility and foaming properties of whey protein suspensions. J. Food Eng. 2008, 86, 281–287. [Google Scholar] [CrossRef]
  104. O′Sullivan, J.; Arellano, M.; Pichot, R.; Norton, I. The effect of ultrasound treatment on the structural, physical and emulsifying properties of dairy proteins. Food Hydrocoll. 2014, 42, 386–396. [Google Scholar] [CrossRef]
  105. Li, H.; Yu, J.; Ahmedna, M.; Goktepe, I. Reduction of major peanut allergens ara h 1 and ara h 2, in roasted peanuts by ultrasound assisted enzymatic treatment. Food Chem. 2013, 141, 762–768. [Google Scholar] [CrossRef]
  106. Zhenxing, L.; Caolimin, L.; Jamil, K. Reduction of allergenic properties of shrimp (penaeus vannamei) allergens by high intensity ultrasound. Eur. Food Res. Technol. 2006, 223, 639–644. [Google Scholar] [CrossRef]
  107. Fonteles, T.V.; Costa, M.G.M.; de Jesus, A.L.T.; de Miranda, M.R.A.; Fernandes, F.A.N.; Rodrigues, S. Power ultrasound processing of cantaloupe melon juice: Effects on quality parameters. Food Res. Int. 2012, 48, 41–48. [Google Scholar] [CrossRef]
  108. Wu, J.; Gamage, T.; Vilkhu, K.; Simons, L.; Mawson, R. Effect of thermosonication on quality improvement of tomato juice. Innov. Food Sci. Emerg. Technol. 2008, 9, 186–195. [Google Scholar] [CrossRef]
  109. Saeeduddin, M.; Abid, M.; Jabbar, S.; Wu, T.; Hashim, M.M.; Awad, F.N.; Hu, B.; Lei, S.; Zeng, X. Quality assessment of pear juice under ultrasound and commercial pasteurization processing conditions. LWT-Food Sci. Technol. 2015, 64, 452–458. [Google Scholar] [CrossRef]
  110. Abid, M.; Jabbar, S.; Hu, B.; Hashim, M.M.; Wu, T.; Lei, S.; Khan, M.A.; Zeng, X. Thermosonication as a potential quality enhancement technique of apple juice. Ultrason. Sonochem. 2014, 21, 984–990. [Google Scholar] [CrossRef]
  111. Scudino, H.; Silva, E.K.; Gomes, A.; Guimarães, J.T.; Cunha, R.L.; Sant′Ana, A.S.; Meireles, M.A.A.; Cruz, A.G. Ultrasound stabilization of raw milk: Microbial and enzymatic inactivation, physicochemical properties and kinetic stability. Ultrason. Sonochem. 2020, 67, 105185. [Google Scholar] [CrossRef] [PubMed]
  112. Shanmugam, A.; Chandrapala, J.; Ashokkumar, M. The effect of ultrasound on the physical and functional properties of skim milk. Innov. Food Sci. Emerg. Technol. 2012, 16, 251–258. [Google Scholar] [CrossRef]
  113. Chen, L.; Chen, J.; Ren, J.; Zhao, M. Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates. J. Agric. Food Chem. 2011, 59, 2600–2609. [Google Scholar] [CrossRef]
  114. Zhou, C.; Ma, H.; Yu, X.; Liu, B.; Yagoub, A.E.-G.A.; Pan, Z. Pretreatment of defatted wheat germ proteins (by-products of flour mill industry) using ultrasonic horn and bath reactors: Effect on structure and preparation of ace-inhibitory peptides. Ultrason. Sonochem. 2013, 20, 1390–1400. [Google Scholar] [CrossRef]
  115. Zhu, Z.; Zhu, W.; Yi, J.; Liu, N.; Cao, Y.; Lu, J.; Decker, E.A.; McClements, D.J. Effects of sonication on the physicochemical and functional properties of walnut protein isolate. Food Res. Int. 2018, 106, 853–861. [Google Scholar] [CrossRef]
  116. Zhang, Z.; Wang, Y.; Jiang, H.; Dai, C.; Xing, Z.; Mintah, B.K.; Dabbour, M.; He, R.; Ma, H. Effect of dual-frequency ultrasound on the formation of lysinoalanine and structural characterization of rice dreg protein isolates. Ultrason. Sonochem. 2020, 67, 105124. [Google Scholar] [CrossRef] [PubMed]
  117. Zhang, H.; Chen, G.; Liu, M.; Mei, X.; Yu, Q.; Kan, J. Effects of multi-frequency ultrasound on physicochemical properties, structural characteristics of gluten protein and the quality of noodle. Ultrason. Sonochem. 2020, 67, 105135. [Google Scholar] [CrossRef]
  118. Chandrapala, J.; Zisu, B.; Palmer, M.; Kentish, S.; Ashokkumar, M. Effects of ultrasound on the thermal and structural characteristics of proteins in reconstituted whey protein concentrate. Ultrason. Sonochem. 2011, 18, 951–957. [Google Scholar] [CrossRef]
  119. Costa, M.G.M.; Fonteles, T.V.; de Jesus, A.L.T.; Almeida, F.D.L.; de Miranda, M.R.A.; Fernandes, F.A.N.; Rodrigues, S. High-intensity ultrasound processing of pineapple juice. Food Bioprocess Technol. 2013, 6, 997–1006. [Google Scholar] [CrossRef]
  120. Jambrak, A.R.; Mason, T.J.; Lelas, V.; Krešić, G. Ultrasonic effect on physicochemical and functional properties of α-lactalbumin. LWT-Food Sci. Technol. 2010, 43, 254–262. [Google Scholar] [CrossRef]
  121. Arzeni, C.; Martínez, K.; Zema, P.; Arias, A.; Pérez, O.; Pilosof, A. Comparative study of high intensity ultrasound effects on food proteins functionality. J. Food Eng. 2012, 108, 463–472. [Google Scholar] [CrossRef]
  122. Hu, H.; Li-Chan, E.C.; Wan, L.; Tian, M.; Pan, S. The effect of high intensity ultrasonic pre-treatment on the properties of soybean protein isolate gel induced by calcium sulfate. Food Hydrocoll. 2013, 32, 303–311. [Google Scholar] [CrossRef]
  123. Jia, J.; Ma, H.; Zhao, W.; Wang, Z.; Tian, W.; Luo, L.; He, R. The use of ultrasound for enzymatic preparation of ace-inhibitory peptides from wheat germ protein. Food Chem. 2010, 119, 336–342. [Google Scholar] [CrossRef]
  124. Nazari, B.; Mohammadifar, M.A.; Shojaee-Aliabadi, S.; Feizollahi, E.; Mirmoghtadaie, L. Effect of ultrasound treatments on functional properties and structure of millet protein concentrate. Ultrason. Sonochem. 2018, 41, 382–388. [Google Scholar] [CrossRef] [Green Version]
  125. Zhang, H.; Claver, I.P.; Zhu, K.-X.; Zhou, H. The effect of ultrasound on the functional properties of wheat gluten. Molecules 2011, 16, 4231–4240. [Google Scholar] [CrossRef] [Green Version]
  126. Xue, F.; Wang, S.; Liu, H.; Li, C. Effects of high-intensity ultrasound treatment on functional properties of plum (pruni domesticae semen) seed protein isolate. J. Sci. Food Agric. 2018, 98, 5690–5699. [Google Scholar] [CrossRef]
  127. Tiwari, B.; Muthukumarappan, K.; O′donnell, C.; Cullen, P. Inactivation kinetics of pectin methylesterase and cloud retention in sonicated orange juice. Innov. Food Sci. Emerg. Technol. 2009, 10, 166–171. [Google Scholar] [CrossRef]
  128. Tzima, K.; Brunton, N.P.; Lyng, J.G.; Frontuto, D.; Rai, D.K. The effect of pulsed electric field as a pre-treatment step in ultrasound assisted extraction of phenolic compounds from fresh rosemary and thyme by-products. Innov. Food Sci. Emerg. Technol. 2021, 69, 102644. [Google Scholar] [CrossRef]
  129. Ostermeier, R.; Hill, K.; Dingis, A.; Töpfl, S.; Jäger, H. Influence of pulsed electric field (PEF) and ultrasound treatment on the frying behavior and quality of potato chips. Innov. Food Sci. Emerg. Technol. 2021, 67, 102553. [Google Scholar] [CrossRef]
Table
Table 1. Changes in the structural properties of various proteins due to applying a pulsed electric field.
Table 1. Changes in the structural properties of various proteins due to applying a pulsed electric field.
FoodsProcessing ConditionsEffect/Influence on ProteinRef.
Horseradish peroxidase5–25 kV/cm; 207–1242 pulses; 1.5 μs pulse widthLoss of α-helix structure by 42% at 22 kV/cm and 87 pulses[46]
Soybean protein isolate0–15 kV; 1–8 μs pulse width; 1–9 ms pulse cycle; electrode gap of 0.292 cmLoss in α-helix and increased random coils and β-sheets. Changes in disulfide bonds and collapse in a hydrophobic core. Strong PEF resulted in the reburial of hydrophobic residues into the core again.[47]
Soybean protein isolate0–50 kV/cm; 40 μs pulse width; treatment time 4.8 ms.Relocation of turns into structured α-helix after 35 kV/cm. Slight increase in anti-parallel β-sheets and reduction in β-sheets content[48]
Hen egg-white lysozyme35 kV/cm; 0–1200 μs time; 2 μs pulse widthInactivation following conventional first-order model. Loss in α-helix and increased random coils and β-sheets along with hydrophobic collapse at earlier stages of inactivation[49]
Soybean trypsin inhibitor0–40 kV/cm; 0–547 μs treatment time; 2 μs pulse widthNo major changes in secondary structure[50]
Egg white protein and β-lactoglobulin12.5 kV/cm; long length pulsesPartial structure modification[51]
Egg ovalbumin20–35 kV/cm for 180 μs and at 35 kV/cm for 60–240 μsHigh-intensity processing resulted in a loss of α-helix and a decrease in surface hydrophobicity[52]
Egg-white protein5–25 kV/cm; pulse width 8 μs; frequency 500 Hz; residence time 90 sIncreased free sulfhydryl groups and total number of sulfhydryl groups decreased. Reduced the α-helix content, while β-sheets increased[53]
Pepsin0–34.2 kV/cm; 23, 28 °CLoss in β-sheets resulting in a loss in activity of the enzyme[54]
Pepsin25.2–35.6 kV/cm; 0–500 μsHydrophobic collapse. Reduction in β-sheets and increased intermolecular hydrophobic interactions and random coils[55]
Whey protein isolate12–20 kV/cm; 10–30 pulses at 0.5 Hz;Reduction in surface hydrophobicity, which can result in structural modifications[56]
Canola protein10–35 kV/cm; pulse width 8 μs; residence time 180 sIncreased voltage and processing time resulted in reduced β-sheets and α-helix. Increased free sulfhydryl groups and reduction in total number of sulfhydryl groups. Increased surface hydrophobicity[57]
Myofibrillar proteins (from PSE like chicken breast0–28 kV/cm; pulse frequency 0–1000 Hz; residence time 180 sModerate PEF application increased solubility and surface hydrophobicity. α-helix increased and β-turns, and random coils reduced with applying PEF intensity[58]
Table
Table 2. Changes in the physicochemical and functional properties of various proteins due to applying a pulsed electric field.
Table 2. Changes in the physicochemical and functional properties of various proteins due to applying a pulsed electric field.
FoodsProcessing ConditionsEffect/Influence on ProteinRef.
Soybean trypsin inhibitor0–40 kV/cm; 0–547 μs treatment time; 2 μs pulse widthDenaturation and aggregation resulted in a reduction of solubility, surface hydrophobicity and free sulfhydryls[50]
Egg white protein and β-lactoglobulin12.5 kV/cm; long length pulsesPartial aggregation and aggregated gel microstructure in EW. Gelation behavior improved in β-lactoglobulin, and it reduced in egg white.[51]
Ovomucin-depleted egg white1.4–1.8 kV/cm; specific energy input of 260–700 kJ/kg; 20 μs pulse widthProtein aggregation at pH 5 and 7, but not at pH 4 and 9. Only lysozyme was responsible for aggregate formation compared to thermal processing[59]
Egg ovalbumin20–35 kV/cm for 180 μs and at 35 kV/cm for 60–240 μsImmunogenic-binding capacity increased for low-intensity processing or high-intensity and short-time processing. Immunogenic-binding capacity decreased for high-intensity processing for >60 μs[52]
Whey protein isolate30–35 kV/cm; 19.2–211 μs; 2 μs pulse width; 30, 60, 65, 70, 75 °CNo effect on protein aggregation, surface hydrophobicity sulfhydryl groups, thermal stability and emulsification properties. Reduction in heat-induced gel strength and increased gelation time[60]
Whey proteins37.6 kV/cm; 50, 100 and 200 pulses of 2 μs at 1 HzNo change in immunoreactivity[61]
Raw milk2–40 kV/cm; 5–35 μs pulse width; 50–1000 Hz; outlet temperature–39–72 °CNo change in color; reduction in conductivity of milk[62,63]
Pepsin25.2–35.6 kV/cm; 0–500 μsIncreased aggregation[55]
Tomato juice35 kV/cm; pulse frequency 50–250 Hz; pulse width 1–7 μs; treatment time 1000 μsApparent viscosity increased with treatment parameters compared to untreated control[64]
Strawberry juice35 kV/cm; pulse frequency 50–250 Hz; pulse width 1–7 μs; treatment time 1000 μsApparent viscosity increased with treatment parameters in the case of monopolar pulses. Bipolar pulses slightly reduced the viscosity[64]
Canola protein10–35 kV/cm; pulse width 8 μs; residence time 180 sImproved solubility, oil-binding capacity, emulsion stability, foamability and water-holding capacity[57]
Almond milk0–28 kV/cm; 40 μs pulse width; 1 kHz frequencyParticle size reduction and stable emulsion. Improved appearance and physical stability at 28 kV/cm[65]
Table
Table 3. Changes in the enzyme activity of various foods due to applying a pulsed electric field. LOX—lipoxygenase; PE—pectinesterase; POD—peroxidase; PPO—polyphenol oxidase; ALP—alkaline phosphatase; HPL—hydroperoxide lyase.
Table 3. Changes in the enzyme activity of various foods due to applying a pulsed electric field. LOX—lipoxygenase; PE—pectinesterase; POD—peroxidase; PPO—polyphenol oxidase; ALP—alkaline phosphatase; HPL—hydroperoxide lyase.
Food/EnzymeProcessing ConditionsEffect/Influence on EnzymeRef.
Horseradish POD5–25 kV/cm; 207–1242 pulses; 1.5 μs pulse widthUp to 37% reduction in activity with increasing pulses and electric field strength[46]
Soybean LOX20–42 kV/cm; 2 μs pulse width; 1036 μs treatment timeMaximum inactivation of 88% at 42 kV/cm when treated for 1036 μs.[66]
Soybean LOX20–40 kV/cm; 25–100 μs; 23, 35, 50 °C85% inactivation at the highest processing conditions[67]
Tomato LOX0–35 kV/cm; 20–70 μs treatment time; 10–50 °CIrreversibly inactivated[68]
Tomato LOX and HPL35 kV/cm; pulse frequency 50–250 Hz; pulse width 1–7 μs; treatment time 1000–2000 μsInactivation resistance of LOX is greater than HPL. 20% and 90% maximum reduction of LOX and HPL, respectively.[69]
Tomato POD35 kV/cm; pulse frequency 50–250 Hz; pulse width 1–7 μs; treatment time 1000–2000 μsInactivation achieved with a minimum pulse frequency of 200 Hz. Maximum inactivation achieved with pulse width > 5.5 μs[70]
Pea LOX2.5–20 kV/cm; 1 μs pulse width; 100–400 pulsesNo inactivation[71]
Watermelon LOX and POD35 kV/cm; 1727 μs treatment time; 4 μs pulse widthLOX more resistant compared to POD inactivation. POD can be completely inactivated, whereas 50% inactivation was observed for LOX for 220 and 250 Hz[72]
POD and PPO in apple juice20–40 kV/cm; 25–100 μs; 23, 35, 50 °CHighest inactivation rates (~70%) at the highest processing values[73]
POD and PPO in grape juice25–35 kV/cm, 600 Hz bipolar pulse width 4 μs, treatment time 5 msComplete inactivation of PPO was achieved, whereas only 50% inactivation was observed for POD[74]
ALP (bovine milk)25–37 kV/cm; 15–60 °C; 2 μs pulse width; treatment time 19.6 μs30–67% inactivation for 25–35 kV/cm at 60 °C[75]
xanthine oxidase (whole milk)20 or 26 kV/cm; pulse width 20 μs; frequency 20 Hz.Inactivation was 7–13% lower compared to thermal processing at 66 °C[76]
Pepsin0–34.2 kV/cm; 23, 28 °CInactivation of pepsin by ~60%[54]
PE in orange juice5–35 kV/cm; 200 Hz; width 4 μs; treatment time 1500 μs20% residual enzyme activity[77]
PE in grapefruit juice20–40 kV/cm; 25–100 μs; 23, 35, 50 °C97% inactivation after treating at 40 kV/cm, 100 μs at 50 °C[78]
Table
Table 4. Changes in the structural properties of various proteins due to applying ultrasonication processing.
Table 4. Changes in the structural properties of various proteins due to applying ultrasonication processing.
Food SourcesProcessing ConditionInfluence on Protein StructureReference
Milk20 kHz frequency, 120 µm
amplitude, 150 W, 55–75.5 °C		α-Lactalbumin and β-lactoglobulin denaturation, up to 81.5% reduction in the size of the fat globule 1[97,98]
β-Lactoglobulin (milk allergen)20 kHz frequency,120 µm amplitude, 135 W/cm2Sonication mostly induced reduction in the β-sheet content while increasing α-helix and/or random coil structure of a protein. Sonication had a minor effect on IgE-binding properties[99]
Black bean protein isolates dispersions (10%, w/v)20 kHz, 150–450 W, 12–24 minDecrease in the α-helix proportion and an increase in β-sheets content in the protein after ultrasonic treatment (300 W, 24 min)[100]
Skim milk20 kHz, 20–40 W, 15–60 minSignificant decrease in the band intensity of β-casein after 15 min sonication. The relative band intensities of β-lac and α-lac (major whey proteins) show a decrease after 30 min sonication. After 45–60 min of sonication, the intensity of the whey proteins is found to be lesser than control. The relative band intensity of κ-casein present in the whey protein (denatured with κ-casein) significantly increased after 30 min treatment at 20 W and 40 W[112]
Soy protein isolate solution (5%, w/v)20 kHz, 0–600 W, 15 min + controlled papain
hydrolysis		Compared to control, US treatment at 400 W combined with a 1.25% degree of hydrolysis can cause a 47.7% reduction in α-helix, 30.4% in β-sheet, and 50% β-turn. A 73.5% increase in the random coil[113]
Defatted wheat germ proteins20 kHz, 0–1800 W for 10 minFree sulfhydryl group and disulfide bonds decreased significantly with increasing the power intensity and sonication time[114]
Walnut protein25 kHz; 15–30 min; 200–600 WReduction in α-helices and increase in β-sheet, random coil and turn components. Increased free sulfhydryl groups compared to control[115]
Faba bean protein20 kHz; 15–30 min; 500/700 WIncreased β-sheet and turn content and reduction in random coil (intermolecular aggregates)[101]
Rice dreg protein isolates20–50 kHz, 15 minUltrasonication altered protein secondary structure by reducing random coil and β-sheet contents, while α-helix and β-turn contents increased[116]
Gluten protein0–40 kHz, 10 min, power density was 67 W/L at 30 °CSonication decreased the α-helix content of all sonicated gluten protein samples while increased the β-sheet and β-turn content, and tryptophan and tyrosine residues were exposed[117]
Almond milk20 kHz, 1–16 min, 450 W at 25 °CUltrasonication increased the ordered structures (α-helix and β-sheet) content[95]
Whey protein solution 2 (cheese)20 kHz, 450 W10% increase in the α-helix component and a 6–9% decrease in the β-sheet and turn components[118]
1 There no change in the property of enzymes when ultrasound was applied without heat generation. 2 Whey protein solution (cheese): The surface hydrophobicity of the proteins increased for up to 5 min of sonication, presumably due to the unfolding of the proteins resulting from minor structural changes. However, the surface hydrophobicity decreased after sonication for more than 5 min, which is a sign of protein aggregation, which in turn protects the hydrophobic regions of the proteins.
Table
Table 5. Changes in the physicochemical and functional properties of various proteins due to applying ultrasonication.
Table 5. Changes in the physicochemical and functional properties of various proteins due to applying ultrasonication.
Food SourcesProcessing ConditionFunctional PropertyReference
Milk600 W at a frequency of 20 kHz and an amplitude of 50%Solubility increased significantly from 35.78% to 88.30% after 5 min. A significant increase in the emulsion stability, surface hydrophobicity and emulsifying activity[97,98]
Pineapple juice19 kHz, 500 W, US intensity was 376 W/cm2, 10 minJuice viscosity reduced by 75% of the initial value[119]
α-Lactalbumin (milk)20 kHz, 15–20 min, 600 WFoam capacities and solubility were improved significantly[120]
Soymilk20 kHz, 450–600 WGreat increase in solubility, specific surface area and emulsion activity index. 16 min US treatment reduced trypsin inhibitor activity of soymilk by 52%[102]
Soy protein isolate (10% w/w solution)20 kHz, 750 W and 20% of amplitude, 20 minNo significant changes in total free sulfhydryl groups and conductivity of protein. Significant increase in surface hydrophobicity (121%), solubility and water holding capacity of protein. A significant reduction in particle size of the protein[121,122]
Wheat germ protein20 kHz, 0–1800 W, 20 minUltrasonic pretreatment caused a 21.0–40.7% increase in ACE-inhibitory activity of defatted wheat germ protein hydrolysate[123]
Whey protein20 kHz, 600 W, 15 min, 43–48 W/cm2Significant increase in the solubility and foamability[103]
Milk protein isolate solution
(0.1–5 wt%)		20 kHz and 95% amplitude, 34 W/cm2, 0–2 minUS treatment reduced the size and hydrodynamic volume of the protein. A significant reduction in the intrinsic viscosity improves the emulsifying activity.[104]
Egg white proteins (10% w/w solution)20 kHz, 750 W and 20% of amplitude, 20 minProtein solutions were not significantly changed after treatment. A significant decrease in solubility[121]
black bean protein isolates20 kHz, 150–450 W, 12–24 minSurface hydrophobicity and protein solubility of protein were enhanced after ultrasonication (300 W, 24 min)[100]
Millet protein concentrate (10% w/w) 120 kHz, 100 W, 5–20 min, 18–74 W/cm2, 20–100% amplitudesSignificant increase in solubility with the processing time and intensity, the highest solubility in 73.95 W/cm2 intensity for 12.5 min. Low ultrasound intensity (18.4 W/cm2) caused an increase in the emulsion activity index and emulsion stability. In contrast, the high ultrasound intensity (73.95 W/cm2) intensity caused a significant decrease in these properties[124]
Ara h 1 and Ara h 2 (Peanut allergen)US–enzyme combination (50 Hz, 1 h, 0–0.30% (w/w) trypsin or α-chymotrypsinProtease digestion greatly increased peanut protein solubility. US–enzyme combination significantly lowered Ara h 1 and Ara h 2 in peanuts. Ultrasound–enzyme combination significantly lowered IgE binding of peanut extract[105]
Shrimp protein30 Hz, 800 W for 1.5 h at 0–50 °CHigh-intensity ultrasound at 50 °C significantly reduced the allergenicity of shrimp (2.2-fold, 2.5-fold lower than control). US treatment caused a 76% reduction in the tropomyosin content[106]
Kiwifruit protein20 kHz, 450 W, 1–16 min at 25 °CUS caused a 50% reduction in the IgE binding capacity of Act d 2. In vitro digestibility of kiwifruit, proteins increased up to 77%[96]
Wheat protein20 kHz; 540, 720, 900 W; 10 min at 25 °CImproved foam capacity, emulsion stability and emulsification properties[125]
Faba bean protein20 kHz; 15–30 min; 500/700 WImproved adsorption dynamics and foamability and reduced the digestibility[101]
Walnut protein25 kHz; 15–30 min; 200–600 WImprovement in solubility and emulsification properties. Reduction in the particle size[115]
Plum seed protein20 kHz; 200–600 WIncreased solubility, emulsifying property and foaming capacity. Improved gel strength and gelling properties. Improved protease accessibility, which can increase digestibility[126]
Whey protein solution (cheese)20 kHz, 450 WSurface hydrophobicity of proteins increased within first 5 min, and it decreased after 5 min[118]
1 A significant increase in solubility with the processing time and intensity, the highest solubility in 73.95 W/cm2 intensity for 12.5 min. The foaming capacity of millet protein solution in lower intensities and times (18.4 W/cm2, 5 min) was significantly lower than the untreated sample. However, its foaming capacity increased significantly as the time of US treatment in high intensities (73.95 W/cm2, 20 min) was prolonged. After 12.5–20 min US treatment at 18.4 W/cm2 intensity, an increase in the emulsion activity index and emulsion stability were observed. 12.5–20 min at 73.95 W/cm2 intensity caused a significant decrease in the emulsion activity index and emulsion stability.
Table
Table 6. Changes in the enzyme activity of various foods due to applying ultrasonication.
Table 6. Changes in the enzyme activity of various foods due to applying ultrasonication.
Food SourcesProcessing ConditionEnzyme ActivityReference
Pineapple juice19 kHz, 500 W, US intensity was 376 W/cm2, 10 min20% reduction in the polyphenol oxidase (PPO) activity (376 W/cm2 and 10 min)[119]
Cantaloupe melon juice19 kHz, 500 W, 376 W/cm2 for 10 minSignificant reduction in peroxidase (POD) and PPO activities[107]
Tomato juice24 kHz, 400 W at amplitudes of 25–75 μm at 60–70 °C90% reduction in the pectin methylesterase activity[108]
Orange juice20 kHz, 0.42–1.05 W/mL, 2–10 minHighest pectin methylesterase (PME) activity inactivation was 62% after 1.05 W/mL sonication for 10 min[127]
Pear juice20 kHz, 750 W, at 25–65 °C for 10 minResidual activities of POD, PME and PPO were 4.3%, 3.25% and 1.91% after sonication at 65 °C for 10 min[109]
Apple juice20 kHz, 5–10 min, 0.30 W/cm3 at 20–60 °CSignificant reduction in enzyme activities of PPO, POD and PME under sonication treatment at 60 °C for 30 min: 63%, 70% and 62%[110]
Raw milk19 kHz, 100–475 W, 1–7 kJ/mLUS treatment promoted microbial and enzymatic inactivation with a temperature below 60 °C[111]
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Vanga, S.K.; Wang, J.; Jayaram, S.; Raghavan, V. Effects of Pulsed Electric Fields and Ultrasound Processing on Proteins and Enzymes: A Review. Processes 2021, 9, 722. https://doi.org/10.3390/pr9040722

AMA Style

Vanga SK, Wang J, Jayaram S, Raghavan V. Effects of Pulsed Electric Fields and Ultrasound Processing on Proteins and Enzymes: A Review. Processes. 2021; 9(4):722. https://doi.org/10.3390/pr9040722

Chicago/Turabian Style

Vanga, Sai Kranthi, Jin Wang, Shesha Jayaram, and Vijaya Raghavan. 2021. "Effects of Pulsed Electric Fields and Ultrasound Processing on Proteins and Enzymes: A Review" Processes 9, no. 4: 722. https://doi.org/10.3390/pr9040722

APA Style

Vanga, S. K., Wang, J., Jayaram, S., & Raghavan, V. (2021). Effects of Pulsed Electric Fields and Ultrasound Processing on Proteins and Enzymes: A Review. Processes, 9(4), 722. https://doi.org/10.3390/pr9040722

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