海獸胃線蟲之海藻糖生成酶活性與低溫相關性研究

Study of the Corelations between Low Temperature and the Trehalose Synthase Activity of Anisakis simplex

曾建仁(Chien-Jen Tseng)

指導教授 ： 施秀惠

國立臺灣大學/生命科學院/動物學研究研究所/碩士(2007年)

https://doi.org/10.6342/NTU.2007.02626

摘要

海獸胃線蟲是海洋經濟魚類常見之蠕蟲類寄生蟲，同時是引起人類海獸胃線蟲症之病原體。由於海獸胃線蟲對冷凍具有抗性，因此食品安全上建議在食用漁產品前需要冷凍足夠時間和溫度以避免被感染。本研究旨在查明海獸胃線蟲經過低溫處理後所產生之抗凍機制，主要探討抗凍蛋白存在與海藻糖生成酶在低溫處理下之活性。使用採集自蘇澳外海鯖魚體內之海獸胃線蟲，以基因體方法和冰親和性純化以了解是否以抗凍蛋白為抗凍機制，結果證明海獸胃線蟲體內並無抗凍蛋白存在。在海藻糖生成酶活性方面，將海獸胃線蟲低溫處理0、4和10 ℃後，所得結果海藻糖生成酶比活性為0.235、0.632和0.819 Unit/mg，對照組16 ℃則為0.151 Unit/mg，在統計上4 ℃及10 ℃和對照組有顯著差異，溫度降低確實使酵素活性增加。0 ℃時和對照組無顯著差異，則推測降溫太快使海獸胃線蟲無法引起抗凍反應表現海藻糖生成酶。而在4 ℃低溫刺激下，12小時海藻糖生成酶活性即有顯著上升，到30小時活性仍有增加。綜和以上的實驗結果，推論海藻糖在海獸胃線蟲中扮演重要的抗凍保護劑，且需要經過低溫刺激才能產生，也支持海獸胃線蟲具有忍耐抗凍能力。

關鍵字

海獸胃線蟲 ； 海獸胃線蟲症 ； 海藻糖生成酶抗凍

並列摘要

Anisakis simplex is a famous helminth parasite of commercial marine fish, which is the pathogen of Anisakiasis. Because A. simplex can survive in freezing temperature, for food safety, it is suggested to keep fishes under freezing temperature long enough to avoid infection before eating sea foods. In this study, the purposes are to investigate antifreeze mechanism of cold treatment, mainly discusses the antifreeze protein existence and the trehalose synthase activity under the low-temperature treatment. This study used the gathering of A. simplex from Spotted Mackerel that were obtained from commercial local fisheries in coastal waters of Su-Ao. The results showed A. simplex does not have the antifreeze protein existence by the genomic analysis and the ice affinity purification. In the aspect of trehalose synthase activity, the activity of trehalose synthase of A. simplex under the low-temperature treatment of 0, 4 and 10 ℃ was 0.235, 0.632 and 0.819 Unit/mg, respectively, and the activity of control group ( treatment of 16 ℃) was 0.151 Unit/mg. In statistics, the result of 4 and 10 ℃ treatment showed remarkable differences with the control group, that is, the decreasing ot temperature indeed caused the increasing of enzyme activity. There is no significant difference between 0 ℃ treatment and control group. It is deduces that, quick temperature decreasing can not induce the anti-freeze response of A. simplex, thus no expression of trehalose synthase. Under 4 ℃ low temperature treatment, the activity of trehalose synthase increased significantly after 12 hours, and remained increased afer 30 hours. In summary, the trehalose may acts as an important cryoprotectant in the A. simplex, but the low temperature stimulation is needed for the production of trehalose. This study supports that A. simplex has the ability of freeze-tolerant.

並列關鍵字

Anisakis simplex ； Anisakiasis ； Trehalose synthase ； Antifreeze

參考文獻

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