本文選題：章魚(yú) + 過(guò)敏原　； 參考：《集美大學(xué)》2012年碩士論文

【摘要】：食物過(guò)敏是人類(lèi)比較常見(jiàn)的一種過(guò)敏性疾病，主要是由食物中的過(guò)敏原引起。人們發(fā)生食物過(guò)敏后，常出現(xiàn)腹瀉、哮喘、嘔吐等癥狀，嚴(yán)重時(shí)會(huì)虛脫、休克，甚至危及生命。水產(chǎn)類(lèi)食物作為重要的過(guò)敏食物之一，由其引起的過(guò)敏反應(yīng)引起了世界人們的關(guān)注，在聯(lián)合國(guó)糧農(nóng)組織公布的八大類(lèi)過(guò)敏食物中，水產(chǎn)品就占了兩大類(lèi)。章魚(yú)作為重要的水產(chǎn)資源，因其肉質(zhì)鮮嫩、味道鮮美，具有很高的營(yíng)養(yǎng)價(jià)值，深受消費(fèi)者喜愛(ài)。隨著章魚(yú)消費(fèi)量的增加，有關(guān)章魚(yú)過(guò)敏的報(bào)道也隨之增多，但是關(guān)于章魚(yú)過(guò)敏原的研究較少，集中于主要過(guò)敏原—原肌球蛋白（TM）的純化及分析，對(duì)于章魚(yú)是否含有其他過(guò)敏原以及如何開(kāi)發(fā)低致敏性章魚(yú)產(chǎn)品等方面尚未見(jiàn)報(bào)道。 本實(shí)驗(yàn)首先對(duì)章魚(yú)TM進(jìn)行分離純化、鑒定及性質(zhì)分析，并制備多克隆動(dòng)物抗體；然后對(duì)章魚(yú)TM分別進(jìn)行超聲聯(lián)合加熱、紫外照射、高溫高壓和美拉德反應(yīng)等加工處理方式；并采用模擬胃腸液消化實(shí)驗(yàn)對(duì)不同處理后TM的消化穩(wěn)定性進(jìn)行分析，進(jìn)一步運(yùn)用水產(chǎn)食物過(guò)敏患者血清對(duì)其致敏性進(jìn)行鑒定；最后，對(duì)章魚(yú)肌肉進(jìn)行熱處理和高溫高壓處理，分析肌肉蛋白和TM的性質(zhì)變化。結(jié)果顯示，純化章魚(yú)TM的分子量為35kDa，與蟹類(lèi)TM具有免疫交叉反應(yīng)，是含糖量為1.3%的糖蛋白，且對(duì)熱和酸堿穩(wěn)定；超聲聯(lián)合加熱和紫外照射對(duì)章魚(yú)TM消化穩(wěn)定性及免疫活性影響不明顯，高溫高壓處理能夠有效降低TM的消化穩(wěn)定性和免疫活性；章魚(yú)TM與葡萄糖發(fā)生美拉德反應(yīng)，隨著反應(yīng)時(shí)間的延長(zhǎng)，其產(chǎn)物的免疫活性逐漸降低，但其消化穩(wěn)定性卻有所增加；對(duì)章魚(yú)肌肉進(jìn)行100℃蒸煮10min和高溫高壓10min處理，發(fā)現(xiàn)兩種處理方式均能夠一定程度地降低章魚(yú)TM的消化穩(wěn)定性，且高溫高壓處理效果更顯著。 本實(shí)驗(yàn)還對(duì)章魚(yú)精氨酸激酶（AK）進(jìn)行分離純化及鑒定，制備多克隆抗體；然后對(duì)其致敏性、熱穩(wěn)定性、pH穩(wěn)定性及消化穩(wěn)定性等理化性質(zhì)進(jìn)行分析；最后將其與甲殼類(lèi)動(dòng)物過(guò)敏原AK進(jìn)行分析比較。結(jié)果顯示，采用水產(chǎn)食物過(guò)敏患者血清對(duì)章魚(yú)肌漿蛋白進(jìn)行免疫反應(yīng)，發(fā)現(xiàn)能與血清發(fā)生特異性反應(yīng)的有50kDa、38kDa和26kDa蛋白組分，其中最主要的是38kDa蛋白組分；進(jìn)一步純化得到該38kDa蛋白組分，應(yīng)用肽指紋圖譜對(duì)其進(jìn)行分析鑒定，確定為章魚(yú)AK；對(duì)章魚(yú)AK的致敏性進(jìn)行分析，確定其為章魚(yú)的一種新型過(guò)敏原，并制備得到了效價(jià)較高的多克隆動(dòng)物抗體；章魚(yú)AK是含糖量為1.1%的糖蛋白，且對(duì)熱和酸堿均不穩(wěn)定，經(jīng)過(guò)不同條件的熱處理和pH處理后，章魚(yú)AK的免疫活性均受到明顯影響；章魚(yú)AK容易被胃蛋白酶、胰蛋白酶及胰凝乳蛋白酶等消化酶降解。對(duì)章魚(yú)AK進(jìn)行分子克隆，得到其全長(zhǎng)基因序列和氨基酸序列，并將章魚(yú)AK與甲殼類(lèi)動(dòng)物過(guò)敏原AK進(jìn)行比較分析。結(jié)果顯示，章魚(yú)AK氨基酸序列比甲殼類(lèi)動(dòng)物AK少8個(gè)氨基酸殘基，且二者序列相似性?xún)H為54%，但二者蛋白三維結(jié)構(gòu)高度相似，且抗原表位預(yù)測(cè)結(jié)果同樣具有一定的相似性；章魚(yú)AK能和甲殼類(lèi)動(dòng)物AK的多克隆動(dòng)物抗體發(fā)生明顯的免疫交叉反應(yīng)；此外，章魚(yú)AK與甲殼類(lèi)動(dòng)物AK具有相似的致敏性，蟹AK能夠抑制章魚(yú)AK與IgE之間的特異性反應(yīng)。 總之，本研究主要是對(duì)章魚(yú)的過(guò)敏原TM和AK進(jìn)行了詳細(xì)分析。首先是對(duì)章魚(yú)的主要過(guò)敏原TM進(jìn)行模擬加工處理，以期篩選出能夠有效降低章魚(yú)TM消化穩(wěn)定性和免疫活性的加工方法；其次是首次證明了AK是章魚(yú)的一種新型過(guò)敏原，，并對(duì)其理化性質(zhì)進(jìn)行進(jìn)一步分析，同時(shí)將其與蝦蟹過(guò)敏原AK進(jìn)行比較，分析二者之間的相似性。這些結(jié)果可望為開(kāi)發(fā)低致敏性章魚(yú)產(chǎn)品提供技術(shù)參考，為章魚(yú)過(guò)敏的診斷和治療提供理論依據(jù)。
[Abstract]:Food allergy is one of the most common allergic diseases in human beings, mainly caused by allergen in food. After food allergy, people often suffer from symptoms of diarrhea, asthma, vomiting and other symptoms, such as deactivation, shock, and even life-threatening. Aquatic food is one of the most important allergic foods, and the allergic reaction caused by it causes the world. Among the eight categories of allergic food published by the United Nations Food and Agriculture Organization, the aquatic products account for two major categories. Octopus, as an important aquatic resource, has high nutritional value because of its delicate meat quality and delicious taste, and is very popular with consumers. With the increase of Octopus consumption, reports on Octopus allergies have increased, but the increase in Octopus allergies is increasing. There are few studies on the allergens of octopus, which are concentrated on the purification and analysis of the main allergens - promyosin (TM). There are no reports on whether Octopus contains other allergens and how to develop low sensitizing Octopus products.
In this experiment, the TM of Octopus was isolated, identified and analyzed, and the polyclonal antibody was prepared. Then the octopus TM was processed by ultrasonic combined heating, ultraviolet radiation, high temperature and high pressure and Maillard reaction, and the digestive stability of TM after different treatments was divided by simulated gastrointestinal digestion experiment. Furthermore, the sensitivities of the patients with aquaculture food allergy were further identified. Finally, the changes in the properties of muscle protein and TM were analyzed by heat treatment and high temperature and pressure treatment of Octopus muscles. The results showed that the molecular weight of the purified octopus TM was 35kDa, and it had a immune cross reaction with the crab TM, and the sugar content was 1.3%. Heat and acid and alkali are stable; ultrasonic combined heating and ultraviolet irradiation have no obvious effects on the digestive stability and immune activity of octopus TM. High temperature and high pressure treatment can effectively reduce the digestive stability and immune activity of TM. The TM and glucose of Octopus have mallard reaction. With the prolonged reaction time, the immune activity of the products gradually decreases, but the effect of the reaction is gradually reduced. The digestive stability was increased, and 100 centigrade steaming 10min and high temperature and high pressure 10min treatment of Octopus muscle found that the two treatments could reduce the digestive stability of the octopus TM to a certain extent, and the effect of high temperature and high pressure treatment was more significant.
The isolation, purification and identification of Octopus arginine kinase (AK), the preparation of polyclonal antibody, and the analysis of its physicochemical properties, such as sensitization, thermal stability, pH stability and digestive stability, were analyzed in the experiment. The results were compared with the allergen AK of the crustaceans. The results showed that the serum of patients with aquatic food allergy was used. The immunoreaction of Octopus sarcoplasmic protein (Octopus sarcoplasmic protein) showed that the protein components of 50kDa, 38kDa and 26kDa could be specific to the serum. The most important component was the 38kDa protein component; the 38kDa protein component was further purified, and the peptide fingerprint was used to identify and identify the octopus AK, and the sensitization of the octopus AK was analyzed. A new type of allergens of Octopus was determined and the polyclonal antibody of high titer was obtained. Octopus AK was a glycoprotein containing 1.1% sugar, and both heat and acid and alkali were not stable. After different conditions of heat treatment and pH treatment, the immunological activity of octopus AK was obviously affected; octopus AK was easy to be pepsin, pancreatic eggs. The molecular cloning of octopus AK was cloned and its full length gene sequence and amino acid sequence were obtained, and the AK of octopus and the allergens of crustaceans were compared and analyzed. The results showed that the AK amino acid sequence of octopus AK was less than 8 amino acid residues in the crustaceans, and the sequence similarity of the two was only 54%. However, the three-dimensional structure of the two proteins is highly similar, and the prediction results of antigen epitopes are similar. Octopus AK can have obvious immunological cross reaction with the polyclonal antibody of crustacean AK; in addition, the octopus AK has similar sensitivities to the crustacean AK, and the crab AK can inhibit the specificity between the octopus AK and IgE. Sexual reaction.
In conclusion, this study is mainly to analyze the allergens of octopus TM and AK in detail. First, the main allergen TM of octopus is simulated and processed in order to screen out the processing methods that can effectively reduce the digestive stability and immune activity of octopus TM. Secondly, it is the first time that AK is a new type of allergens of octopus. Further analysis was carried out, and compared with the allergen AK of shrimp and crab, the similarity between the two was analyzed. These results may provide a technical reference for the development of low sensitizing Octopus products and provide a theoretical basis for the diagnosis and treatment of Octopus allergies.
【學(xué)位授予單位】：集美大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：S917.4;R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 馬之明;徐實(shí)懷;陳怡飚;賈曉平;吳文杰;;真蛸肌肉營(yíng)養(yǎng)成分及氨基酸含量分析[J];安徽農(nóng)業(yè)科學(xué);2011年02期

2 焦海峰,尤仲杰,竺俊全,王一農(nóng);嘉庚蛸對(duì)溫度、鹽度的耐受性試驗(yàn)[J];水產(chǎn)科學(xué);2004年09期

3 胡燕,黎海芪;兒童食物過(guò)敏的診斷方法[J];中國(guó)兒童保健雜志;2000年03期

4 周曉秋;于廣成;王U

本文編號(hào)：1982036