低致敏食品制备技术及其工业化应用研究进展

食物过敏是联合国粮农组织和世界卫生组织认定的全球性食品安全问题之一。在食品加工多元化的背景下,食物过敏患者要完全避免过敏原十分困难,研发低致敏食品对食物过敏患者的安全膳食至关重要。总结了低致敏食品制备技术的加工技术原理;以蒸煮、微波和烘烤为主的热加工技术通过加热诱导蛋白质变性的方式破坏致敏性构象性表位;高压、脉冲电场、脉冲光、低温等离子体、辐照和超声等非热加工技术可以通过过敏原蛋白结构修饰、多肽链断裂、新化学键的产生等方式直接破坏致敏性表位;酶水解、酶交联、糖基化、微生物发酵等其他加工方法则通过改变蛋白质构象或将蛋白质与糖类物质结合,破坏或隐藏过敏原致敏性表位。另外,对工业化低致敏蛋白配料的加工方法和生产现状进行了阐述分析。基于酶法水解的部分水解乳蛋白和深度水解乳蛋白已经可以工业化生产,其他消减食物致敏性的方法以及其他低致敏蛋白配料值得进一步研究。希望可以为工业化生产低致敏食品提供参考。     

Potential Utility of High‐Pressure Processing to Address the Risk of Food Allergen Concerns

High‐pressure processing (HPP) technology is a novel, nonthermal processing technology for food. This special processing method can inactivate microorganisms and enzymes in food at room temperature using ultra‐high pressures of above 100 MPa, while the original flavor and nutritional value of the food are maintained, with an extended refrigerated shelf‐life of the food in distribution. In recent years, because of the rising prevalence of food allergies, many researchers have actively sought processing methods that reduce the allergenicity of food allergens. This study describes the effects of the current HPP technology on allergen activity. Our main goal was to provide an overview of the current research achievements of the application of HPP to eliminate the allergenicity of various foods, including legumes, grains, seafood, meat, dairy products, fruits, and vegetables. In addition, the processing parameters, principles, and mechanisms of HPP for allergen destruction are discussed, such as the induction of protein denaturation, the change in protein conformation, allergen removal using the high‐pressure extraction technology, and the promotion of enzymatic hydrolysis to alter the sensitization of the allergens. In the future, the application of HPP technology as a pretreatment step for raw food materials may contribute to the development of food products with low or no allergenic ingredients, which then can effectively reduce the concern for consumers with allergies, reduce the risk of mistaken ingestion, and reduce the overall incidence of allergic reactions from food.     

Effect of roasting history and buffer composition on peanut protein extraction efficiency

Peanut is a major allergenic food. Undeclared peanut (allergens) from mis-formulation or contamination during food processing pose a potential risk for sensitized individuals and must be avoided. Reliable detection and quantification methods for food allergens are necessary in order to ensure compliance with food labelling and to improve consumer protection. The extraction of proteins from allergenic foods and complex food products is an important step in any allergen detection method. In this study, the protein extraction efficiency of various buffers prepared in-house and some extraction buffers included in some commercial allergen enzyme-linked immunosorbent assay (ELISA) test kits for peanut determination in food products were tested. In addition, the effect of roasting history on the extractability of peanut protein was investigated by the biuret and the bicinchoninic acid (BCA) assays. Elevated roasting temperatures in food processing were found to have a major impact on protein extraction efficiency by reducing protein yields of oil and dry roasted peanuts by 50-75% and 75-80%, respectively, compared with the raw material. Extraction buffers operating in the higher pH range (pH 8-11) showed best yields.     

Advanced DNA- and Protein-based Methods for the Detection and Investigation of Food Allergens

Currently, food allergies are an important health concern worldwide. The presence of undeclared allergenic ingredients or the presence of traces of allergens due to contamination during food processing poses a great health risk to sensitized individuals. Therefore, reliable analytical methods are required to detect and identify allergenic ingredients in food products. The present review addresses the recent developments regarding the application of DNA- and protein-based methods for the detection of allergenic ingredients in foods. The fitness-for-purpose of reviewed methodology will be discussed, and future trends will be highlighted. Special attention will be given to the evaluation of the potential of newly developed and promising technologies that can improve the detection and identification of allergenic ingredients in foods, such as the use of biosensors and/or nanomaterials to improve detection limits, specificity, ease of use, or to reduce the time of analysis. Such rapid food allergen test methods are required to facilitate the reliable detection of allergenic ingredients by control laboratories, to give the food industry the means to easily determine whether its product has been subjected to cross-contamination and, simultaneously, to identify how and when this cross-contamination occurred.     

从过敏原危害评估食物过敏风险

随着全球食物过敏发生率的逐年升高,食物过敏性疾病已成为人们日益关注的食品安全和公共卫生问题.本文介绍了食物过敏原特性、致敏机制、临床表现、常见致敏食物及主要暴露来源等内容,概括了食物过敏原闽值及参考剂量的制定方法和最新调整情况,并总结了目前国际上公认的几种食物过敏原的风险评估方法,以期对下一步开展我国人群食物过敏风险评...     

Whey allergens: Influence of nonthermal processing treatments and their detection methods

Whey and its components are recognized as value-added ingredients in infant formulas, beverages, sports nutritious foods, and other food products. Whey offers opportunities for the food industrial sector to develop functional foods with potential health benefits due to its unique physiological and functional attributes. Despite all the above importance, the consumption of whey protein (WP) can trigger hypersensitive reactions and is a constant threat for sensitive individuals. Although avoiding such food products is the most successful approach, there is still a chance of incorrect labeling and cross-contamination during food processing. As whey allergens in food products are cross-reactive, the phenomenon of homologous milk proteins of various species may escalate to a more serious problem. In this review, nonthermal processing technologies used to prevent and eliminate WP allergies are presented and discussed in detail. These processing technologies can either enhance or mitigate the impact of potential allergenicity. Therefore, the development of highly precise analytical technologies to detect and quantify the existence of whey allergens is of considerable importance. The present review is an attempt to cover all the updated approaches used for the detection of whey allergens in processed food products. Immunological and DNA-based assays are generally used for detecting allergenic proteins in processed food products. In addition, mass spectrometry is also employed as a preliminary technique for detection. We also highlighted the latest improvements in allergen detection toward biosensing strategies particularly immunosensors and aptasensors.     

Application of real-time PCR for tree nut allergen detection in processed foods

Abstract

Currently, food allergies are an important health concern worldwide. The presence of undeclared allergenic ingredients or the presence of traces of allergens due to accidental contamination during food processing poses a great health risk to sensitized individuals. Therefore, reliable analytical methods are required to detect and identify allergenic ingredients in food products. Real-time PCR allowed a specific and accurate amplification of allergen sequences. Some processing methods could induce the fragmentation and/or degradation of genomic DNA and some studies have been performed to analyze the effect of processing on the detection of different targets, as thermal treatment, with and without applying pressure. In this review, we give an updated overview of the applications of real-time PCR for the detection of allergens of tree nut in processed food products. The different variables that contribute to the performance of PCR methodology for allergen detection are also review and discussed.     

Development of a Biological Assay to Determine the Allergenic Potential of Foods

Food allergies represent a risk for many people in industrialized countries. Unrecognizable allergenic proteins of foodstuffs may be present as ingredients that are not labeled or as unknown cross-contamination. Such hidden allergens can cause severe reactions in allergics, even at minute quantities, sometimes with fatal outcome. For the verification of the presence of allergenic food constituents, analytical methods such as ELISA and PCR have been developed. However, these tests cannot measure allergenic potential. For this reason, a test system that measures the biological activity of allergens has been developed. It is based on the cellular mechanisms of the type I allergy. Rat basophilic leukemia cells (RBL-2H3) were transfected with the genes of the human high affinity receptor for IgE. The resulting cell line expressed the human receptor α-chain and could bind allergenspecific IgE from allergic subjects, in contrast to the parent cell line. After cross-linking of receptor-bound, allergen-specific human IgE by allergens, the cells released measurable inflammatory mediators. These cells were used for the analysis of a variety of allergen extracts, including extracts prepared from foods containing allergenic hazelnut and peanut. The comparative validation with existing ELISA and PCR for hazelnut and peanut demonstrated similar sensitivity and specificity. The established cell line will be a novel tool in the detection of allergens in complex mixtures, especially to address the issue of their allergenic potential, which cannot be accomplished by classical analytical methods. This will add valuable information about the allergenic potential of food constituents to the risk assessment of foods.     

Use of lysozyme as an indicator of protein cross-contact in fresh-cut vegetables via wash waters

To evaluate risks linked to cross-contact of allergenic proteins in food production sites via carry-over, it is necessary to gain more insight in the behavior of proteins during processing. A typical case example of cross-contact is related to the re-use of wash water in the fresh-cut vegetable processing industry. In this study, the carry-over of allergenic proteins via wash water was quantified by applying an allergen-indicator, lysozyme. The adsorption of the allergen to the fresh-cut vegetables could be characterized by a Langmuir adsorption isotherm. From the adsorption characteristics, it was observed that the carry-over was significantly stronger on carrots compared to lettuce and leek. This was moreover observed from the mass balance which illustrated that the total amount of lysozyme transferred from the wash water to the fresh-cut vegetables was in average 77.4, 25.1 and 22.2% for respectively carrots, lettuce and leek. From a deterministic risk assessment, it could be concluded that allergenic proteins can be transferred via wash water to fresh-cut vegetables in the next production batch in such quantities that they pose a risk towards allergic consumers. The proposed methodology enables the food industry to validate designed preventive measures in the framework of their allergen management.     

Characterization of plant food allergens: An overview on physicochemical and immunological techniques

Allergy to plant‐derived foods is a highly complex disorder with clinical manifestations ranging from mild oral, gastrointestinal, and cutaneous symptoms to life‐threatening systemic conditions. This heterogeneity in clinical manifestations has been attributed to different properties of allergenic molecules. Based on this fact, symptom elicitors were grouped into class I and pollinosis‐associated class II food allergens, but clear distinction is rather ambiguous. Moreover, mechanisms underlying food sensitization are not fully understood yet, and food allergy management most often relies on patient's compliance to avoid suspected foods. Therefore, recent efforts aim at the investigation of plant food allergies at the molecular level. This review provides an overview on currently available techniques for allergen characterization and discusses their application for investigation of plant food allergens. Data obtained by an array of physicochemical analyses, such as allergen structure, integrity, aggregation, and stability, need to be linked to results from immunological methods at the level of IgE and T‐cell reactivity. Such knowledge allows the development of computational algorithms to predict allergenicity of novel foods being introduced by biotechnological industry. Furthermore, molecular characterization is an indispensable tool for molecule‐based diagnosis and future development of safer patient‐tailored specific immunotherapy in plant food allergy.     

Considerations on Developing Criteria to Determine Whether a Food Allergen is of Public Health Importance

Food allergy is the result of a particular type of immune response against one or more food components, usually proteins. The food allergy reactions that are the focus of regulatory measures are mediated by antibodies of the IgE class. Importantly, food allergy is a two-step process. The first step, the sensitisation phase, consists of an immune response resulting in production of IgE antibodies specific for the allergen. The second step, the provocation phase, is the triggering of a symptomatic allergic reaction as a result of exposure to the allergen after sensitisation has been established. For reasons not well understood, a majority of sensitised individuals never will experience clinical reactions. Determining the number of sensitised individuals (test positives) in a population will therefore grossly overestimate the prevalence of food allergy. The term ‘allergy’ should be used only if clinical reactions occur. For triggering a food allergic reaction in a sensitised individual, only the ‘acute’ food intake and not the intake over time is of importance, and food allergy in this aspect resembles acute poisoning. Because of the cultural, agricultural, economic and nutritional importance of the foods, society accepts that a small fraction of the population develops allergies to traditional food products. Prevention of allergic reactions is then sought by means of education and by labelling of the most important allergenic foods. However, some 200 allergenic foods have been described, but only a small minority of these appears to be of importance in terms of frequency and severity of reactions triggered in the population. When it comes to management of the allergens, however, there is a lack of clear criteria at two levels: at the level of defining what documentation should be required to enter a proposed new food allergen into one of the large allergen databases, as well as at the level of determining which food allergens are of sufficient public health importance to require special regulatory attention in terms of labelling. With the aim of increasing transparency and predictability of decision-making processes and obtaining more consistency between labelling of different allergens as well as between labelling of allergens in different food regulatory jurisdictions, ILSI Europe has taken an initiative to establish clear criteria and a framework to determine the public health importance of a given food allergen. These criteria will consist of factors relating to food properties, population factors, and exposure factors.     

Why are some proteins allergenic? Implications for biotechnology

In recent years, a number of agricultural crops have been developed with recombinant DNA technology. Because the transferred genes code for proteins that are ordinarily not present in these particular foods, there is concern about the potential allergenicity of these new crop varieties. Foods contain many proteins; however, only a small fraction are allergens. Although the structural properties of proteins that cause allergic reactions have not been characterized completely, known food allergens in general have molecular weights between 10 and 70 kDa, stimulate the immune response (induce the production of allergen‐specific IgE), and are stable molecules that are resistant to processing, cooking, and digestion. Although any type of food is potentially allergenic, the majority of food allergies are caused by a small group of foods (cows’ milk, nuts, legumes, eggs, seafood). Cross‐reactivities occur within a given food group and between foods and seemingly unrelated proteins.

Even though most transgenic foods are considered safe, biotechnological manipulation can affect crop allergenicity. The safety evaluation of transgenic foods is relatively easy when the allergenicity of the gene sources are known. The recombinant food can be assayed using traditional in vitro inhibition assays. Recently, reduced allergen content of biotechnologically altered rice was shown. In contrast, increased allergenicity was demonstrated in transgenic soybeans after a methionine‐ and cystine‐rich protein from Brazil nuts, identified as a major Brazil nut allergen, was expressed in soybean to increase its content of sulfur‐rich amino acids. The most difficult issue regarding transgenic food allergenicity is the effect of transfer of proteins of unknown allergenicity. The challenge is to determine whether these proteins are allergenic as there is no generally accepted, established, definitive procedure to define or predict a protein's allergenicity. Comparing the structures of the transferred protein with known allergens and allergen epitopes could be one approach. Additionally, Th‐2 cell stimulation, the production of IL‐4, and IgE antibody induction in animal models may help to evaluate the potential allergenicity of a protein.

In conclusion, there is no evidence that recombinant proteins in newly developed foods are more allergenic than traditional proteins. The evidence suggests that the vast majority of these proteins will be completely safe for the consumer. The concern is that if a few transgenic foods cause serious allergic reactions, this could undermine the public's confidence in such products. It is essential that proper guidelines are established and tests are developed to assure that this will not occur.     

Methods for allergen analysis in food: a review

Food allergies represent an important health problem in industrialized countries. Undeclared allergens as contaminants in food products pose a major risk for sensitized persons. A proposal to amend the European Food Labelling Directive requires that all ingredients intentionally added to food products will have to be included on the label. Reliable detection and quantification methods for food allergens are necessary to ensure compliance with food labelling and to improve consumer protection. Methods available so far are based on protein or DNA detection. This review presents an up-to-date picture of the characteristics of the major food allergens and collects published methods for the determination of food allergens or the presence of potentially allergenic constituents in food products. A summary of the current availability of commercial allergen detection kits is given. One part of the paper describes various methods that have been generally employed in the detection of allergens in food; their advantages and drawbacks are discussed in brief. The main part of this review, however, focuses on specific food allergens and appropriate methods for their detection in food products. Special emphasis is given to allergenic foods explicitly mentioned in the Amendment to the European Food Labelling Directive that pose a potential risk for allergic individuals, namely celery, cereals containing gluten (including wheat, rye and barley) crustaceans, eggs, fish, peanuts, soybeans, milk and dairy products, mustard, tree-nuts, sesame seeds, and sulphite at concentrations of at least 10 mg kg^-1. Sulphites, however, are not discussed.     

基于蛋白质和DNA的食品过敏原检测技术的研究进展

目前,食品过敏问题已成为全球性的健康问题。在食品加工过程中产生的食品过敏成分或微量过敏原对敏感机体都是巨大的健康威胁。因此,可靠的分析方法是鉴别和检测食品中过敏成分所必需的。该文综述了基于蛋白质和脱氧核糖核酸(DNA)的食品过敏原检测技术的应用及发展,并展望了其未来发展趋势。     

食品过敏原检测与评价技术研究进展

过敏原生物活性包括过敏原性即引起致敏个体发生过敏症的活性和致敏原性即引起人群致敏的危险性两个方面。随着转基因作物及相应食品的大量出现，评价和检测食品过敏原生物活性日益受到重视。迄今，食品过敏原性的主要检测方法有：皮肤试验、双盲安慰剂对照激发试验、血清IgE检测和组胺释放试验等；对食品致敏原性还没有建立可靠的评价和监测技术，FAO/WHO采纳的分级评价策略包括血清学测定、过敏原分子结构和序列同源性比较及胃肠液消化稳定性评价等。本文对食品过敏原生物学活性的评价与检测技术研究进展进行综述。     

食品过敏原的安全管理

本文综述了重大的相对性食品安全问题食品过敏原的特点和安全管理措施。主要阐述了食品过敏原的产生机制,主要类别和特点,检测与评价技术的发展并建议从加强标签管理、建立专业数据库、开发新产品、严格规范加工过程以及如何防范和治疗等几个方面着手来努力实现对这类食品安全问题的管理。     

Assessment of allergic potential of (novel) foods

In safety assessment of Novel Foods such as functional foods, allergy is a special issue on which particular emphasis has been placed. The reason for such concern is that incidence of food allergies is constantly and rapidly increasing. The severity of the reported incidents and the number of foods incriminated are also on the rise. The outstanding challenge is to understand what makes a common innocuous protein or peptide behave as an allergen for some groups of people, or why it may suddenly or progressively become a much more potent allergen than usual. It is therefore necessary to consider the risks of creating or unmasking new immunoreactive structures, or of overexposure to already reactive substances, as a result of new food-production and processing technologies. No test such as the use of animal models, the analysis of structure, function and physico-chemical properties is as yet available to evaluate or predict the allergenicity of a “novel” protein in a wholly reliable and objective manner. No indication has yet suggested that novel foods, and particularly recombinant proteins or genetically modified foods, would be more (or less) allergenic than the corresponding conventional foods. No particular structure can be described as being solely and intrinsically allergenic. The predictive approaches to determining the allergenic potential of NFs should therefore be subject to case-by-case critical appraisal allied to mandatory implementation of monitoring of the potential postmarketing impact of these new foodstuffs on public health.     

蟹类主要过敏原及其消减技术研究进展

蟹是引起食源性过敏的主要食品之一,蟹类过敏原是引起蟹过敏的根源。近年来,过敏原性质的研究及分离纯化技术备受关注,已发现蟹的主要过敏原包括原肌球蛋白、精氨酸激酶、血蓝蛋白等。利用食品加工技术降低蟹致敏性的研究也日益增多,以热加工法、辐射技术、酶处理法和超高压法等为代表的蟹致敏活性消减技术取得了新进展。本文就蟹类主要过敏原的种类、致敏蛋白的制备、蟹与其他甲壳类的交叉过敏性以及蟹类过敏原消减方法等进行综述,以期为蟹过敏的诊断预防和低致敏产品开发提供参考。     

Impact of irradiation and thermal processing on the immunochemical detection of milk and egg allergens in foods

There is little information available on the antigenicity of allergens in foods subjected to gamma irradiation and thermal processes. The objective of the study was to evaluate the effects of gamma irradiation and thermal processing on the recovery of milk and egg allergens in foods prepared using irradiated wheat flour incurred with these allergens. Bread, boiled pasta and extruded cereal were selected as food matrices for the study.Allergen detection was performed using Surface Plasmon Resonance (SPR) and commercially available enzyme linked immunosorbent assay (ELISA) kits. Protein structure was characterized using differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR) spectroscopy, and circular dichroism (CD) spectroscopy. The ELISA kits performed well in detecting allergens in the unprocessed flours with recoveries ranging from 63–120%. The results for dough and uncooked pasta were similar to those of unprocessed flours. Recoveries for boiled pasta were significantly (p ≤ 0.05) reduced. The lowest allergen recoveries were obtained in bread samples. Allergen recoveries in gamma irradiated samples depended on the irradiation dose, the type of ELISA kit used, the tested matrix, and the processing method. FTIR and CD results showed that among the allergens casein was the most thermostable followed by ovomucoid. β-Lg and ovalbumin showed thermal sensitivity. In conclusion, gamma irradiation may affect the antigenicity of allergenic food residues at low levels depending on the food matrices and the processing methods.     

Oral tolerance induction does not resolve gastrointestinal inflammation in a mouse model of food allergy

Scope : Oral immunotherapy (OIT) involving continuous oral administration of allergenic foods has gained attention as a therapy for food allergies. To study the influence of oral administration of allergenic foods on gastrointestinal symptoms including inflammation, we established a mouse model of food‐induced intestinal allergy. Methods and results : BALB/c mice were fed an egg white (EW) diet containing ovalbumin (OVA, a major EW allergen) after intraperitoneal sensitisation with OVA and Alum. The mice on the EW diet for one wk presented gastrointestinal symptoms (i.e. weight loss and soft stools) and inflammation in the small intestines (i.e. duodenum, jejunum and ileum). Further continuous EW diet resolved the weight loss but not the soft stools. Splenic CD4⁺ T‐cells of EW diet‐fed mice on the continuous diet showed less proliferation and cytokine production compared with those of control mice, suggesting tolerance induction by the diet. The continuous EW diet reduced levels of OVA‐specific IgE antibodies, but significantly aggravated the inflammation in the jejunum. Conclusion : Our mouse model would be useful to investigate inflammatory and regulatory mechanisms in food‐induced intestinal allergies. Our results suggest potential gastrointestinal inflammation in patients undergoing OIT as continuous administration of allergenic foods, even though the therapy may induce clinical tolerance.