南美白对虾主要过敏原原肌球蛋白的低过敏性处理方法研究

以南美白对虾原肌球蛋白(Tropomyosin,Tm)为研究对象,采用酶解法、超声结合酶解法消减3种虾制品中的Tm。首先,以菠萝蛋白酶水解富集的Tm样品通过优化酶活与底物质量比、反应时间和反应温度,建立了能有效消减Tm的酶解方法;同时对比了酶解、超声结合酶解法分别对虾仁、蝴蝶虾仁和虾糜3种制品中的Tm过敏原性变化情况。Tm致敏动物模型的抗血清ELISA结果显示,单纯酶解和超声结合酶解处理的虾仁其过敏原性无显著性差异(P0.05);经超声结合菠萝蛋白酶酶解处理后的蝴蝶虾仁样品其过敏原性降低了21.05%;而虾糜样品中,单纯的酶解或超声结合酶解法与对照相比均有显著性差异,其过敏原性分别减少了30.70%和33.33%。综上所述,作者建立的酶解法可有效消减Tm的过敏原性,该酶解法以及超声结合酶解法在低过敏原性蝴蝶虾仁和虾糜制品的生产领域具有较高的应用可行性。     

Reduction of allergenic properties of shrimp (Penaeus Vannamei) allergens by high intensity ultrasound

The tropomyosin fraction of shrimp proteins is potentially responsible for an allergic reaction in individuals with a genetic predisposition to allergy. However, there are no efficient and safe methods to reduce its allergenicity. High-intensity ultrasound is known to change the structure of proteins. The aim of this study was to assess the effect of high-intensity ultrasound on the IgE-binding capacity of shrimp protein extracts (PE) and of major shrimp allergen Pen a 1(tropomyosin). Peeled shrimp (Penaeus vannamei) muscle was sealed in a plastic bag and treated with high-intensity ultrasound (30 Hz, 800 W) for 1.5 h at 0 and 50 °C, respectively; others were treated with boiling water for 15 min. Then PE and Pen a 1 were made with treated shrimp muscle. The allergenicity of PE and purified allergen from different-treated shrimp were analyzed by the enzyme allergosorbent test (EAST) and competitive inhibition ELISA (Ci-ELISA) using sera of 15 shrimp-allergic patients. PE from high-intensity ultrasonic treatment at 50 °C (treated 2) shrimp was 2.2-fold, 2.5-fold lower respectively than that of untreated shrimp and high-intensity ultrasonic treatment at 0 °C (treated 1) shrimp. While PE from heated (treated 3) shrimp was 1.2-fold lower than that of untreated shrimp. The (Concentration of the different-treated sample that inhibit 50% of the IgE binding to coated untreated sample) IC₅₀ ratios of Pen a 1 prepared from treated 2 shrimp, treated 1 shrimp and treated 3 shrimp to untreated shrimp were 1/15, 1/1.1 and 1/1, respectively. The results suggest that high-intensity ultrasound at 50 °C may reduce the allergenicity of shrimp.     

高强度超声对凡纳滨对虾蛋白结构和功能特性的影响

为探究高强度超声对凡纳滨对虾蛋白结构和功能特性的影响，本实验采用300 W的超声功率、20 kHz的超声频率和382 W/cm2的超声功率强度对对虾肉全蛋白匀浆分别超声0、5、15、25、35 min，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和酶联免疫吸附试验检测高强度超声处理后对虾原肌球蛋白含量的变化，发现高强度超声波处理可以有效降低对虾肌肉蛋白的致敏性；利用傅里叶变换红外光谱分析高强度超声处理后对虾蛋白的二级结构变化，结果显示高强度超声波处理使蛋白质二级结构从无规卷曲向β-转角和β-折叠转化；同时，通过测定巯基含量发现高强度超声处理后对虾肌肉蛋白巯基含量增加，这也有助于形成稳定的三维空间网络结构；此外，对高强度超声处理后对虾的理化和功能特性进行分析，结果表明超声波处理后样品的平均粒径明显降低，对虾肌肉蛋白的总抗氧化能力、游离氨基酸含量以及蛋白体外模拟消化率都明显提高。由此可见，高强度超声处理在降低对虾蛋白致敏性的同时，能够改变其结构和功能特性，提高其潜在的营养价值，本实验可为凡纳滨对虾的非热加工和精深加工提供一定的参考。     

Altering allergenicity of cow's milk by food processing for applications in infant formula

Cow's milk-based infant formulas have a long tradition in infant nutrition, although some infants are unable to use them due to presence of several known allergens. Various processing methods have been identified capable of reducing cow's milk protein allergenicity including thermal and non-thermal methods and their combinations. Heat treatment and enzymatic hydrolysis have been in production of hypoallergenic infant formulas. However, modulation of allergenic epitopes depends on the extent of heat treatment applied, which consequently may also reduce a nutritional value of these proteins. In addition, enzymatic hydrolysis may not target allergenic epitopes thus allergenicity may persist; however released peptides may have detrimental impact on taste and functional properties of final products. Modulation of allergenicity of milk proteins appears to require a concerted effort to minimize detrimental effects as clinical studies conducted on commercial hypoallergenic formulas demonstrated persistence of allergic symptoms. This article covers traditional and novel processing methods and their impact on reduction of cow's milk allergenicity in milk-based infant formulas.     

市售低值贝类加工食品中原肌球蛋白致敏性评估及其消减技术

本实验评估了市售低值贝类及其加工食品中主要致敏原——原肌球蛋白（tropomyosin,TM）的致敏性,并对贝类可食部位中TM致敏性的消减技术进行研究。使用双抗体夹心酶联免疫吸附法分析市售低值贝类及其冻制类、干制类、烟熏类和蒸煮类加工食品中TM含量;选取具有较高致敏性的贝类作为研究对象,探究传统加工技术（干制、蒸煮、高温高压、速冻）和新兴加工技术（高静水压、超声波、糖基化）对TM致敏性的消减效果。结果表明,市售贝类及其加工食品中普遍存在致敏性,TM含量范围在18.94～22.51 ng/g之间。在4 种新鲜贝类中,蚬子的TM含量最高,为（22.51±1.06）ng/g。与未处理的蚬子样品相比,干制、蒸煮和糖基化处理的蚬子中TM致敏性（以TM含量表征）没有显著差异（P＞0.05）,而高温高压、速冻、高静水压和超声波处理均能显著降低蚬子中TM的致敏性（P＜0.05）,其中,以超声波（500 W、30 min）和高温高压处理的消减效果较好,致敏性消减率分别增加了32.4%和26.9%。本研究结果为低致敏性贝类加工食品的开发提供了一定的理论基础和技术支撑。     

The effect of high-pressure treatment on functional components of shrimp (Litopenaeus vannamei) cephalothorax

High-pressure (HP) technology has been applied to extend the shelf life of shrimps by inhibiting enzymes with PPO activity or microorganisms. However, there is very little information on its effect on relevant compounds from a nutritional or functional point of view, such as fatty acids, α-tocopherol, astaxanthin, and hemocyanin, which constitutes the main objective of the present work. Shrimp cephalothoraxes were HP processed at 200, 400, or 600 MPa/18 °C/15 min or three consecutive 5 min cycles. It was found that hemocyanin was partially denatured at pressures up to 400 MPa, resulting in lower PPO activity, and it was totally denatured at 600 MPa, although 20% residual PPO activity remained. Astaxanthin, α-tocopherol, and total antioxidant activity were stable whichever HP treatment was applied, whereas 600 MPa caused a slight reduction of eicosapentaenoic acid (C20:5n3, EPA) and docosahexaenoic acid (C22:6n3, DHA). Despite this reduction, the ω-6/ω-3 fatty acids ratio was very low (1).Industrial relevanceShrimps are high-value fishery products with a very short shelf life under refrigeration, mainly because of microbial growth and development of melanosis. Thermal treatment is effective for extending shelf life, but it affects the nutritional quality of shrimps through degradation of bioactives such as polyunsaturated fatty acids, tocopherols, or astaxanthin, which are mainly located in the cephalothorax. High pressure is a non-thermal processing technology that has been proved to extend shrimp shelf life, but very little information can be found on its effect on the above-mentioned compounds as well as on the melanosis-inducing hemocyanin. Such basic knowledge is very important for industrial application of high-pressure technology to extend the shelf life of shrimps.     

蛋白溶出与变性结合消减虾仁致敏性

以南美白对虾虾仁为原料,采用高压下蛋白溶出与变性相结合的方法消减虾仁致敏性。在室温下采用不 同压力（0.1～900.0 MPa）处理虾仁,确定适合虾仁蛋白溶出和变性的压力条件,并以虾仁蛋白致敏性的消减效果 为指标确定虾仁的处理条件。结果显示,在200.0 MPa下保压处理30 min,有利于虾仁蛋白的溶出;当压力大于等 于500.0 MPa时,可引起虾仁蛋白的变性;当压力在600.0 MPa时,虾仁蛋白的致敏性最小。将虾仁在200.0 MPa下 处理30 min后,再用600.0 MPa处理30 min,虾仁的致敏性消减率大于80%。因此,高压处理可消减虾仁的致敏性, 采用适合蛋白溶出与变性相结合的压力处理,能够提高虾仁致敏性消减效果。     

Protein hydrolysates of salted duck egg white improve the quality of Jinga Shrimp (Metapenaeus affinis)

The influence of protein hydrolysates of salt duck egg white (PHSDEW) hydrolysed by acid protease on the quality of Jinga Shrimp (Metapenaeus affinis) was analysed. Shrimp immersed in 5% PHSDEW with degree of hydrolysis (DH 3%) had the highest values of cooking yield, water retention, moisture and texture with the lowest cooking loss when compared to which immersed in 0.4% NaCl, 3.5% mixed phosphates and other PHSDEW with different DH (1%, 2% and 5%) (P < 0.05). This treatment made muscle fibres retain the most appropriate compact and regular microstructure and improved the thermal stability of shrimp protein as well (P < 0.05). SDS‐PAGE protein patterns revealed shrimp protein would dissolve out during immersion, while only could PHSDEW infiltrate into shrimp muscle. It suggested PHSDEW would be developed as a new type of water retention agent replaced phosphate used in shrimp or other seafoods, which will decrease the waste of salt duck egg white.     

Effect of high-pressure treatment on in-vitro digestibility of β-lactoglobulin

The effect of high-pressure (HP)-treatment on β-lactoglobulin (β-Lg) was investigated using in-vitro pepsin digestion under simulated gastric conditions. HP-treatment of β-Lg at 400 MPa for 10 min only slightly increased its subsequent hydrolysis by pepsin. However, higher pressure treatments (600 and 800 MPa) resulted in rapid digestion of β-Lg. After these higher pressure treatments, β-Lg disappeared in less than 1 min of pepsin incubation as determined by SDS-PAGE analysis. Mass spectrometry analysis of the digestion products at corresponding incubation times revealed rapid and progressive degradation of β-Lg. Most (> 90%) of the peptide products following pepsin digestion of HP-treated β-Lg were less than 1500 Da in size. Peptide products from pepsin digestion were identified and mapped to β-strand regions (Leu₃₂–Leu₅₄ and Phe₈₂–Leu₁₀₄) and to the N- and C-terminals regions (Leu₁–Leu₁₀ and Ser₁₅₀–Leu₁₅₆) of β-Lg. While these regions corresponded to known IgE epitopes of β-Lg, the predominant peptides resulting from 60 s of incubation were short (7–10 residues) in length. These results demonstrate that HP-treatment increased the digestibility of β-Lg and represents a promising processing technology for reducing the allergenicity of known allergens in a wide variety of food materials.Industrial relevanceHigh-pressure treatment is widely used to enhance the functional attributes of food proteins. The potential for enhanced nutritional value of β-Lg was also demonstrated here by its increased digestibility. High-pressure treatment followed by incubation with proteases may represent a method for the commercial production of bioactive peptides such as inhibitors of angiotensin converting enzyme. More importantly, high-pressure-induced unfolding of milk proteins may reduce their allergenicity. Unfolded proteins are less likely to become agents of immunological sensitization because they are more readily hydrolyzed. Thus high-pressure treatment applied to food ingredients such as whey protein isolate may contribute to the development of hypoallergenic foods.     

Shrimp processing residue as an alternative ingredient for new product development

Shrimp residues were dried (65 °C), grounded, and posteriorly used as an ingredient to the production of ‘spiced shrimp flour’ and ‘shrimp flavoured biscuits’. Both products were packed in modified atmosphere (100% N₂) and stored for 180 days (25 °C) for shelf life evaluation. The centesimal composition, physicochemical, microbiological and sensory analyses were carried out in triplicate. The microbiological analysis (residue and shrimp flour) was within the limits of the legislation, confirming the hygienic–sanitary care during processing. The protein content was the most outstanding (40.13% for the spiced shrimp flour and 20.52% for the shrimp flavoured biscuits). The microbiological evaluation for the ‘spiced shrimp flour’ and ‘shrimp flavoured biscuits’ was below the legal limit, and both products were accepted by sensory analysis. The shelf life evaluation demonstrated stability for 6 months. Thus, we concluded that the Litopenaeus vannamei shrimp residue is a promising ingredient in the food industry.     

Penetration of Sodium Tripolyphosphate into Fresh and Prefrozen Peeled and Deveined Shrimp

The distribution of sodium tripolyphosphate (STP) in peeled and deveined shrimp tails after treatment was investigated using P³² labeled STP. Fresh and frozen brown (Penaeus aztecus) and fresh and frozen white (Penaeus setiferus) shrimp were used in the study. Shrimp were treated in solutions of either 0.5%, 1%, 5%, or 10% STP that had been prepared using P³² labeled STP diluted with “cold” STP. The duration of treatment was either 20 sec, 1 min, 5 min, or 20 min. When shrimp were treated in 0.5% and 1% STP solutions, a phosphate concentration gradient was evident in the shrimp muscle. After such treatments, STP was shown to accumulate on the surface of the muscle preventing further STP uptake. At higher STP concentrations (5% and 10% solutions) prolonged time treatments overcame the concentration gradient as STP became equally distributed through the shrimp muscle (4.8 mm). No difference in the STP penetration mechanism between fresh and prefrozen treated shrimo was evident.     

Ultrasound‐induced changes in physical and functional properties of whey proteins

Use of high‐intensity ultrasound to modify certain functional properties of whey proteins is an alternative to traditional method in food industry. Whey protein isolate (WPI) solutions were treated with an ultrasound probe (20 kHz) at different intensities (20% or 30% amplitude) and durations (10 or 20 min). Results showed that ultrasound treatment changed physical and several functional properties of whey proteins including decreased particle size (from 190.4 nm to 138.0 nm), increased surface hydrophobicity (from 5.13 × 10⁵ to 5.77 × 10⁵), free sulphydryl groups (from 52.64 μmol SH g^?1 to 53.64–58.77 μmol SH g^?1), solubility (from 74.95% to 89.70%), emulsion activity index (from 3.18 m² g^?1 to 3.59–5.32 m² g^?1) and emulsion stability index (from 62.26 min to 71.44–104.83 min), and changed viscosity (from 5.51 mPa.s to 4.81–5.64 mPa.s). Therefore, we conclude that high‐intensity ultrasound can be potentially applied to whey proteins to improve its specific functions during food processing.     

热加工与非热加工技术对水产品致敏性的影响研究进展

我国作为水产养殖和消费总量第一大国,水产品引发的过敏问题越来越受到关注,近些年来,因食用水产品而导致的过敏事件日益增多。该研究综述了近年来热加工技术（蒸、煮、高温压力）、非热加工技术（超高压、低温等离子体、超声波和辐照）以及两种加工技术联用对水产品过敏原消减的研究,指出热加工技术主要通过使蛋白质变性来消减过敏原的致敏性,非热加工技术则通过掩盖或破坏过敏原抗原表位来消减过敏原致敏性,为低致敏性水产品开发提供了重要基础和技术参考。不断探究过敏原诱导过敏反应发生的机理,加快推进低致敏性水产品加工技术在实际生产中的应用,有利于控制和降低水产品过敏所带来的风险。     

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。     

Peanut Allergy: Characteristics and Approaches for Mitigation

Peanut allergy has garnered significant attention because of the high sensitization rate, increase in allergy, and severity of the reaction. Sufficiently reliable therapies and efficient mitigating techniques to combat peanut allergy are still lacking. Current management relies on avoiding peanuts and nuts and seeds with homologous proteins, although adverse events mostly occur with accidental ingestion. There is a need for hypoallergenic peanut products to protect sensitized individuals and perhaps serve as immunotherapeutic products. Alongside traditional practices of thermal and chemical treatment, novel processing approaches such as high‐pressure processing, pulsed ultraviolet light, high‐intensity ultrasound, irradiation, and pulsed electric field have been performed toward reducing the immunoreactivity of peanut. Covalent and noncovalent chemical modifications to proteins also have the tendency to alter peanut allergenicity. Enzymatic hydrolysis seems to be the most advantageous technique in diminishing the allergenic potential of peanut. Furthermore, the combined processing approach (hurdle technologies) such as enzymatic hydrolysis followed by, or in conjunction with, roasting, high pressure and heat, ultrasound with enzymatic treatment, or germination have shown a significant reduction of peanut immunoreactivity and may emerge as useful techniques in reducing the allergenicity of peanut and other foods. This study represents our current knowledge about the alterations in allergenic properties of peanut via different processing mechanisms as well as evaluating its future potential, geographical based data on increasing sensitization, clinical relevance, eliciting dose, and current management of peanut allergy. Furthermore, the molecular characteristics and clinical relevance of peanut allergens have been discussed.     

加工对鸡蛋过敏原的影响

卵类黏蛋白、卵白蛋白、卵转铁蛋白和溶菌酶是鸡蛋中的主要过敏原,适当的物理、化学和生物加工可以降低它们的致敏性。其中物理法生产的低致敏性蛋制品可供鸡蛋轻微过敏人群食用或作为免疫治疗药物;而化学和生物法制备的低过敏鸡蛋制品存在安全风险,它能否应用于生产,还有待进一步研究。另外,各种加工方法可以影响过敏原蛋白二硫键及高级结构甚至一级结构,从而导致该蛋白的致敏性发生变化。总之,加工对鸡蛋过敏原结构和致敏性的影响仍然是值得深入探索的科学问题,对指导生产和研发低致敏性或无致敏性蛋制品具有重要作用。     

高压结合酶法消减南美白对虾虾仁致敏性

以南美白对虾虾仁为原料,研究高压结合酶法对虾仁过敏原致敏性的消减作用。将南美白对虾去头去尾去壳去肠线后,采用超高压法和高压结合酶法消减其致敏性,用间接酶联免疫吸附法检测致敏性消减效果,确定消减条件。结果表明：采用超高压法处理,在压力100 MPa、温度25 ℃、保压时间15 min的条件下,虾仁过敏原的致敏性降低了67.09%,且与处理过程中蛋白溶出量有关;采用高压结合酶法处理,在盐水质量浓度1 g/100 mL、酶与虾仁质量比1∶130、压力450 MPa、温度40 ℃、保压时间55 min的条件下,虾仁过敏原的致敏性降低了86.58%。由此可见,高压处理对虾仁过敏原的致敏性有消减作用,高压结合酶法的消减效果更好。     

Impact of high intensity pulsed electric field on antioxidant properties and quality parameters of a fruit juice-soymilk beverage in chilled storage

The effects of high-intensity pulsed electric fields (HIPEF) treatment (35 kV/cm, 4 μs bipolar pulses at 200 Hz for 800 or 1400 μs) on the microbial stability, quality parameters and antioxidant properties of a fruit juice-soymilk (FJ-SM) beverage along the storage time at 4 °C were compared to those obtained by thermal pasteurization (90 °C, 60 s). HIPEF processing for 800 μs ensured the microbial stability of the beverage during 31 days; however, longer microbial shelf-life (56 days) was achieved by increasing the treatment time to 1400 μs or by applying a thermal treatment. Peroxidase and lipoxygenase of HIPEF treated beverages were inactivated by 17.5-29% and 34-39%, respectively; whereas thermal treatment achieved 100% and 51%. During the storage, vitamin C content and antioxidant capacity depleted with time, and they were higher in FJ-SM beverages processed by HIPEF than in those thermally treated. Instead, total phenolic content of beverages did not present significant changes over the time, and it was higher in the 1400 μs-HIPEF treated beverages. In general, color, soluble solids, pH, and acidity values were not significantly affected by the processing treatments. Beverage viscosity increased over time, regardless of the treatment applied. Hence, application of HIPEF may be a good alternative treatment to thermal processing in order to ensure microbiological stability, high nutritional values and fresh-like characteristics of FJ-SM beverages.