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。     

花生过敏原蛋白分离纯化方法研究进展

花生中已确定的过敏原蛋白包括Ara h 1～Ara h 11 11种。本文详细介绍花生中主要过敏原蛋白(Ara h 1、Ara h 2、Ara h 3/4、Ara h 6)以及非主要过敏原蛋白(Ara h 7～Ara h 11)的分离纯化方法研究进展。花生过敏原蛋白的分离纯化方法包括硫酸铵沉淀法、柱层析法、电泳法。其中硫酸铵沉淀法主要用于粗提纯化过程,而柱层析法则主要用于花生过敏原蛋白的精制,它包括离子交换层析、凝胶过滤层析、亲和层析、疏水相互作用层析、高效液相色谱。目前离子交换层析和凝胶过滤层析在花生过敏原蛋白分离纯化中应用最为广泛,而电泳法则仅见应用于Ara h 7及油质蛋白(Ara h 10、Ara h 11)的分离纯化。     

加工导致的花生过敏原蛋白结构变化及其对性质的影响

花生是世界粮农组织(FAO)认证的八大过敏食物之一,花生可导致严重的过敏反应,甚至危及生命。由于花生作为食物配料有广泛的应用,人们对花生过敏越来越关注。本文对加工后花生过敏原蛋白结构变化与性质变化之间的关系研究进行了综述,为探索加工定向改变花生过敏原蛋白的结构和性质提供方向。      

加工方式对花生致敏性的影响及其致敏性评价研究进展

花生蛋白是一种理想的食品加工原料,同时也是一种致死率较高的食物过敏原,严重危害人类健康。本文主要综述食品加工方式对花生过敏蛋白致敏性的影响,以及体内、体外致敏性评价方法,为低致敏或无致敏花生制品的开发提供参考和指导。     

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。     

花生过敏原及其检测方法研究进展

花生过敏可导致某些人群严重的食品安全问题。过敏患者只能通过避免食用含有花生过敏原成分的食物来避免过敏。但是,食品在生产加工、储存、运输、销售过程中有可能被过敏原污染。因此,确定各类加工食品中是否含有花生过敏原成分,对于预防食用者发生花生过敏反应具有重要意义。花生中已确定的过敏原蛋白有13种(Ara h 1~Ara h 13)。本文综述了花生中过敏原蛋白的结构信息,当前流行的提取方法,以及各种定性定量的检测方法,总结了各种方法的优缺点。同时对建立一种具有特异性强、灵敏度高、定量准确的花生致敏蛋白检测方法的发展趋势进行了展望。     

加工对食物过敏蛋白致敏性影响的研究进展

食物过敏已成为世界关注的重大公共卫生和食品安全问题。文章综述近年来国内外有关加工处理(热加工和非热加工)对食物过敏蛋白致敏性的影响:加工方式、处理强度等均会影响过敏蛋白的分子特性,从而导致其或增敏、或脱敏、或不变;加工虽不能完全消除过敏原的致敏潜力,但可通过加工方式和加工参数的选择使其致敏潜力最小化。通过选择合适的食品加工方式控制食物过敏原,在不改变食物营养价值的条件下,获得脱敏性食物,满足食物易敏人群的正常饮食需求,为消费者提供安全食品,是现代食品工业的重要任务之一。     

芝麻过敏原及其致敏性消减技术研究进展

芝麻是新的“8大类”过敏食物之一,芝麻过敏反应由于其潜在危害性及其日益上升的发病率而越来越引起全世界的普遍重视,因此研究芝麻致敏性消减技术在保障食品安全方面具有重要意义。概述了芝麻主要过敏原(Ses i 1～Ses i 7)的结构和免疫特性,及其致敏性消减技术原理和研究进展;已报道的芝麻过敏原有7种,属于2S白蛋白、7S类豌豆球蛋白、油质蛋白和11S球蛋白,其中11S球蛋白和2S白蛋白是主要的过敏原蛋白。利用热加工技术消减芝麻致敏性,主要通过煮沸、微波、烘焙等工艺引起过敏原的解聚、变性进而破坏致敏表位;高压、辐照、发酵、酶解等非热加工技术通过氢键、疏水键等化学键的变化、多肽链的断裂引起蛋白结构变化,从而掩盖或直接降解致敏表位。另外,结合其他食品过敏原,对芝麻过敏原的潜在消减技术,包括脉冲电场、冷等离子体、超声波、脉冲光、糖基化改性和复合加工技术等进行了阐述分析。未来需要进一步研究芝麻不同过敏原的致敏表位、不同加工工艺对过敏原结构及致敏性影响、开展血清学、细胞和动物模型等致敏性评价等,以明确芝麻致敏性消减的主要机制,以期为生产低敏或脱敏芝麻产品奠定理论依据与科学指导。     

压电免疫传感技术快速筛检花生中过敏原

目的采用压电免疫传感技术快速筛检花生过敏原。方法对花生主要过敏蛋白进行分离纯化,免疫新西兰大白兔获得花生抗体,对其的效价,半数抑制浓度等值进行测定。运用自组装的压电免疫传感器包被花生抗体,制作标准曲线,确定灵敏度,回收率及重现性。结果分离纯化花生过敏原蛋白纯度为90.7%,最适的花生包被抗原浓度为0.1μg/mL,自组装的压电传感器检测花生过敏原浓度在1~316μg/mL范围内具有较好的线性关系,灵敏度为0.1μg/mL,回收率介于92.3%～113.4%之间,重现性变异系数为6.04%。结论本项目建立的压电免疫传感技术检测花生过敏原与目前较多的光学分析相比灵敏度更高,实现了快速、在线检测,成本低,仪器简单,具有广阔的应用空间。     

时间分辨免疫荧光法测定食物中花生过敏原成分

目的:建立双抗体夹心时间分辨荧光免疫法[Time-resolved Fluoroimmunoassay(TRFIA)]测定食物中花生过敏原蛋白成分,为进出口食品中花生过敏原成分检测和由花生导致的食物过敏性疾病的预防提供技术基础。方法:提取花生蛋白,免疫小鼠制备抗花生总蛋白的多克隆抗体,用该抗体包被酶标板,并用生物素标记该多抗作为捕捉抗体,Eu3+标记的链酶亲和素和以β-二酮体为主的增强液等试剂,建立双抗夹心TRFIA方法,检测该方法的灵敏度,同时用于13种食品中花生过敏原蛋白成分检测。结果:初步地研制出双抗体夹心TRFIA法检测食品中花生过敏原蛋白成分,具有特异性,和其最低检出限为0.1ng/mL,标准曲线在0.1～160ng/mL范围内线性良好;11种食品检测结果与食物过敏原标签标注内容相符,而2种标示不详的食品检测结果均呈现阳性。     

Investigation on sequential extraction of peanut allergens for subsequent analysis by ELISA and 2D gel electrophoresis

A two-step sequential extraction method of peanut proteins was proposed with the aim to investigate the protein composition and allergen content of peanut samples. The extraction procedure reported is fully compatible with subsequent analysis by enzyme-linked immunosorbent assays (ELISA) as well as 2D gel electrophoresis (2D PAGE). This sequential extraction method was used to study three different peanut varieties and three different types of food processing. Peanuts were analysed for total protein content and the extraction efficiency of raw and processed peanuts was determined. The total protein content of the three peanut varieties was found to be comparable, but their extraction efficiency varies. The peanut extracts were characterised by employing three different ELISA test kits specific to either the allergens Ara h 1 or Ara h 2, or to soluble peanut proteins. The content of both Ara h 1 and Ara h 2 differed in the raw peanut extracts of the three varieties. However, thermal processing resulted in much larger changes in detectability. Blanching significantly increases the detectability of Ara h 2, whereas Ara h 1 detection remains almost unchanged. After roasting a clear decrease of detectability was observed for both Ara h 1 and Ara h 2, although the effect is more severe for Ara h 1. 2D PAGE was employed to compare the protein profiles and abundances of peanut extracts. Statistically relevant differences were observed for the two different protein fractions obtained by using the described method, showing the relevance of this two-step sequential extraction method.     

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.     

花生红衣多酚类物质的制备及其抗氧化活性研究进展

花生红衣是花生加工过程中的廉价副产品,含有丰富的酚类物质,具有良好的抗氧化活性。本文在对花生红衣多酚文献分析统计的基础上,综述了花生红衣多酚类物质水提法、有机溶剂提取法、超声波辅助提取法、微波辅助提取法、酶辅助提取法和闪式提取法等制备方法,有机溶剂萃取法、大孔树脂分离法和膜分离法等分离纯化方法及组分分析方法,抗氧化活性评价方法以及其在食品工业中的应用,并对其研究中的重点和存在的问题进行了总结和展望。     

Allergenicity of peanut allergens and its dependence on the structure

Food allergies are a global food safety problem. Peanut allergies are common due, in part, to their popular utilization in the food industry. Peanut allergy is typically an immunoglobulin E-mediated reaction, and peanuts contain 17 allergens belonging to different families in peanut. In this review, we first introduce the mechanisms and management of peanut allergy, followed by the basic structures of associated allergens. Subsequently, we summarize methods of epitope localization for peanut allergens. These methods can be instrumental in speeding up the discovery of allergenicity-dependent structures. Many attempts have been made to decrease the allergenicity of peanuts. The structures of hypoallergens, which are manufactured during processing, were analyzed to strengthen the desensitization process and allergen immunotherapy. The identification of conformational epitopes is the bottleneck in both peanut and food allergies. Further, the identification and modification of such epitopes will lead to improved strategies for managing and preventing peanut allergy. Combining traditional wet chemistry research with structure simulation studies will help in the epitopes’ localization.     

花生致敏性评估方法及食品加工对其致敏性影响的研究进展

花生是“8大类”过敏原食物之一，因其诱发严重的临床症状及日益上升的发病率而受到广泛关注。文章概述了花生过敏原(Ara h 1-18)的结构特性、免疫特性及体内和体外评估方法，总结了不同食品加工对花生致敏性的研究进展，并展望了未来食品加工技术对消减花生致敏性的研究方向。     

花生蛋白亚基结构与性质研究进展

花生蛋白是一种营养与功能兼具的优良植物蛋白,具有广阔应用前景,花生蛋白结构与功能性质研究是一直以来的研究热点。主要总结了花生蛋白亚基组成与分类、结构与性质,对花生蛋白亚基分离纯化、结构特点、构效关系等方面研究进展进行了详细论述,同时指出目前该研究领域中存在的问题,并对未来的研究重点进行展望,为高品质花生蛋白产品的进一步开发利用提供参考。     

原料及热处理对花生致敏性的影响

花生过敏严重影响健康。简述花生中主要过敏原的一般特征,详细介绍了原料及热风干燥、蒸煮、煎炸和焙烤等热处理对花生致敏性的影响,为研究和开发无过敏或低过敏性花生制品提供了一定的科学依据。     

Mitigation of peanut allergenic reactivity by combined processing: Pressured heating and enzymatic hydrolysis

Among food allergens, peanut is one of the most critical. This study evaluates peanut allergenic features after the combination of heat, pressure, and enzymatic digestion under sonication, by immunodetection using serum IgE of sensitized patients and mass-spectroscopy. In the studied population, there was a predominance of patients with sensitization to Ara h 9 (nsLTP) followed by sensitization to seed storage proteins (Sprot, Ara h 1, 2, 3, and 6). The Sprot sensitized patients showed higher reactivity. The enzyme E5 was efficient for inducing protein fragmentation and allergenic reactivity reduction when it was used combined with pressured heating treatments such as autoclave and Controlled Instantaneous Depressurization (DIC). Only a few Ara h 1 and Ara h 3 peptides were identified after enzymatic digestion of DIC peanut samples. The combination of pressured heating treatments and enzymatic hydrolysis was the most efficient method to strongly mitigate or even eliminate the allergenic potential of peanut. Our findings set a possibility for a group of patients in which their allergy could be treated with a processed less-allergenic peanut and consequently less risky, more easy and quicker desensitization treatment.Industrial relevanceThe findings identify innovative thermal, pressure and enzymatic processing conditions highly effective to mitigate or even abolish the allergenic potency of peanut, which may be relevant for consumers, clinicians, regulatory agencies and the food industry. The applications of processed peanut with reduced IgE binding potency for tolerance induction might be a convenient strategy.