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花生过敏原蛋白分离纯化方法研究进展 总被引:2,自引:0,他引:2
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。 相似文献
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Arah2蛋白是花生的主要过敏原蛋白之一,为获得高纯度的花生Arah2蛋白,以新鲜花生为原料,通过蛋白浸提、DEAE-Sepharose Fast Flow阴离子交换层析,十二烷基磺酸钠—聚丙烯酰胺凝胶电泳(SDS—PAGE)回收等方法分离得到目的蛋白,并运用基质辅助激光解吸/电离飞行时间质谱(MALDI—TOF/MS)对其进行鉴定。结果表明:该方法可得到纯度较高的纯化蛋白;经质谱鉴定后确定该蛋白为Arah2蛋白,两个同种异型物分子量分别为18.5 kD(Arah2.01)和20.1kD(Arah2.02);层析柱分离和SDS—PAGE电泳回收的得率分别为31.4%和18.6%。 相似文献
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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。 相似文献
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食品加工对过敏原活性的影响 总被引:1,自引:1,他引:1
近年来, 食物过敏性疾病的发病率明显上升, 已成为影响人类健康最常见的全球性疾病之一。本文对食品加工中常用的几种方法对特定食品过敏原活性的影响进行了综述, 这些加工方法包括热处理、高压处理、研磨、辐照、碱水解、梅拉德反应、酶解、微生物发酵和基因工程等, 并对存在的问题进行了分析。 相似文献
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目的 探究酪蛋白糖巨肽与花生过敏原相互作用并降低其免疫反应性的潜力。方法 通过蛋白-蛋白分子对接技术探讨酪蛋白糖巨肽(casein glycomacropeptides, CGMP)与Ara h1、Ara h2是否有相互作用的潜力。进一步通过混合水浴加热制备CGMP与花生蛋白的混合溶液(mixed solution of casein glycomacropeptides and peanut proteins, MCGP),建立MCGP致敏、花生蛋白激发的BALB/c小鼠模型,研究MCGP对花生过敏反应的影响。最后使用圆二色谱法研究酪蛋白糖巨肽与Ara h1、Ara h2的相互作用力及对其结构的影响。结果 CGMP与Ara h1、Ara h2间存在次级键(盐桥、氢键、范德华力),部分作用于过敏原表位;MCGP致敏组血清中的花生蛋白特异性免疫球蛋白E(Specific immunoglobulin E, sIgE)、sIgG1、sIgG2a含量显著下降,白介素-4(interleukin-4, IL-4)、IL-5、转化生长因子-β(transforming growth factor-β, TGF-β)、组胺水平显著下降,肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)水平显著升高, MCGP中Ara h2的α-螺旋与β-折叠的比例改变。结论 CGMP能够改变Ara h2的结构,遮蔽花生过敏原表位,抑制sIgE、sIgG结合Ara h1、Ara h2,降低部分花生过敏原的免疫反应性。 相似文献
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花生多肽的提取、分离及纯化研究 总被引:1,自引:0,他引:1
研究了从冷榨花生饼中提取、分离和纯化花生多肽。用碱溶酸沉法从冷榨花生饼中提取花生蛋白,用AS1.398中性蛋白酶水解花生蛋白得到花生多肽粗品,超滤截取相对分子量小于5kD和3 kD的两部分,冷冻干燥后用Sephadex G-25柱层析分析分别得到2个和1个活性峰,在三甲基甘氨酸SDS-聚丙烯酰胺凝胶电泳上显示单一条带的纯品,用5 kD超滤得到的2个峰的相对分子量在6.5 kD和2 kD左右,而用3 kD超滤得到的1个峰的分子量在2 kD左右,Sephadex G-25结合超滤的方法可以获得较纯的花生多肽。 相似文献
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时间分辨免疫荧光法测定食物中花生过敏原成分 总被引:1,自引:0,他引:1
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。 相似文献
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Junjun Liu Wenbin Zhang Pengfei Li Zhumao Jiang Ruijin Yang 《International Journal of Food Science & Technology》2020,55(9):3203-3214
A two-stage microfiltration and ultrafiltration (MF/UF) for peanut protein recovery and water recycling from aqueous extraction processing (AEP) were investigated. Peanut protein aggregates and alkaline water were obtained using flat and wound MF/UF and alkaline water recycled in AEP. With this approach, most of the wastewater could be recycled to produce reusable water to solve wastewater problems in aqueous oil extraction processing from oilseeds. The result showed that a little oil and protein were lost (1.21% and 4.35%, respectively) during the recycling of permeate in AEP, which was still acceptable. Peanut protein aggregations (PPAs) were characterised by analysis with Fourier-transform infrared spectroscopy (FTIR), sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE), high-performance gel filtration chromatography (HPGFC) and binding force analysis. Solubility analysis suggested that PPA had molecular structures with more hydrophobic groups on the surface than in the peanut protein isolate (PPI). In vitro, PPA was digested by both pepsin and trypsin more than PPI. FTIR profiles demonstrated there were more β-sheet and less α-helix in PPA, which means much more aggregation. In addition, binding force analysis showed hydrophobic interaction was the major force that restricted dissolution of PPA and disulfide bonds were of second importance, which provided a possible physical treatment opportunity for improving the solubility of PPA. 相似文献
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Qin Geng Ying Zhang Min Song Xiaoya Zhou Yu Tang Zhihua Wu Hongbing Chen 《Comprehensive Reviews in Food Science and Food Safety》2023,22(2):1058-1081
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. 相似文献
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介绍了蛋白质、蛋白酶以及短肽类研究的一些传统方法和新兴方法。主要对分离、提纯、鉴定及分析方法的使用条件、适用范围、操作步骤、各自特点和应用价值作了简要阐述,并对有关方法进行了比较与评价。 相似文献