台灣市售常見單棘魨之蛋白質圖譜生物多樣性分析法建立

Studies on biological diversity of Monacanthidae in Taiwan by using protein map

趙彥清(Chao Yan-Ching)

指導教授 ： 謝承紘

亞洲大學/健康學院/保健營養生技學系碩士班/碩士(2009年)

摘要

台灣發生之水產品食物中毒事件中，較嚴重者多數由食用有毒魨科魚類或其相關製品所引起的。目前於市場上所販售之香魚片或罐頭，大多數以魨科魚類為原料，因此市售魚類加工品之魚種標示及其食用安全之重要性，是不容忽視的。
本篇論文乃利用SDS-PAGE蛋白質電泳技術分析台灣產主要五種新鮮單棘魨魚種包括杜氏刺鼻單棘魨、長尾革單棘魨、單角革單棘魨、單棘魨及冠鱗單棘魨之水溶性、鹽溶性蛋白質及SDS、Urea萃取之總蛋白質圖譜，比較其應用於魚種鑑別之適切性。結果獲知水溶性、鹽溶性蛋白質圖譜及利用SDS萃取之總蛋白質圖譜其低分子量區(≦30 KD)可區分此五種魚種，而以Urea萃取之總蛋白質圖譜較不易區分此五種魚種。本實驗亦同時探討比較市售快速檢測螢光套組所萃取蛋白質圖譜與傳統SDS-PAGE技術之蛋白質圖譜之優劣，結果顯示市售快速檢測不易區分此五種魚種，故不適用於魨科魚種鑑別之用。
其次，本研究同時亦探討此檢測技術應用於魨科魚類加工品之魚種鑑別的可行性。將五種單棘魨魚種以不同加熱條件處理 (100℃加熱5至30分鐘；121℃加熱5至30分鐘，製成香魚片)，結果顯示加熱100℃之水溶性低分子量區蛋白質之特異電泳帶可鑑別魚種，但鹽溶性低分子量區蛋白質之特異電泳帶較不易作為鑑別魚種指標，而SDS總蛋白質之特異電泳帶隨加熱時間增加並無太大變化，故亦可利用作為魚種判別，但Urea萃取之總蛋白質圖譜中種特異電泳帶會隨加熱時間增加而明顯裂解，故不適宜用來判別魚種。加熱121℃之水溶性、鹽溶性蛋白質及SDS、Urea萃取之總蛋白質種特異電泳帶皆隨加熱時間增加而明顯裂解，故蛋白質技術及圖譜分析法不適宜應用於經高溫加熱後之魚肉加工品。進一步應用此技術於自製香魚片、市售香魚片與市售罐頭，結果得知自製及市售香魚片其蛋白質圖譜種特異電泳帶仍存在，但市售罐頭可能因加熱時間及溫度過久且過高，而易造成蛋白質嚴重變性，其蛋白質圖譜種特異電泳帶已裂解不見。
因此，本篇論文證實水溶性、鹽溶性蛋白質圖譜及以SDS萃取之總蛋白質圖譜鑑別法適宜作為新鮮單棘魨魚類魚種判別之檢測方法。另外，此方法應用於加工製品時，可初步鑑別輕度熱加工之市售香魚片，而無法應用於高度熱加工之罐頭食品之魚種檢測。

關鍵字

蛋白質圖譜 ； 魚種鑑別

並列摘要

Many marine food poisoning incidents due to ingestion of toxic filefishes or its derivative products have occasionally occurred in Taiwan. However, dried dressed fish fillets and canned fish meats are sold in the market and it’s major material is commonly filefishes. Therefore, it is an important issue for products labeling and public food safety.
In this study, we utilized the protein technology (SDS-PAGE) to compare the varieties of the sarcoplasmic protein, myofibrillar protein and Urea, SDS extraction of total protein profile in the 5 species of fresh filefish, including Cantherhines dumerilii, Aluterus scriptus, Aluterus monoceros, Monacanthus chinensis and Stephanolepis cirrhifer. Then, these protein profiles were employed to identify the fish species. It indicated that each different fish species have different species-specific bands in the molecular weight region below 30 KD in their sarcoplasmic, myofibrillar protein and SDS extraction of total protein electrophoresis patterns but not Urea extraction of total protein. We also compared the quality of protein species-specific bands extracted between Fluorescent SPRINT NEXT GELTM kit and SDS-PAGE. The result has shown that use of Fluorescent SPRINT NEXT GELTM kit to identify fish species is ineffective. Therefore, this method does not suitable for the identification of filefish species.
Additionally, in order to understand the application of the aforementioned techniques in processing products of filefish, we have tried the different heating processes (100C heating for 5 to 30 minutes; 121C heating for 5 to 30 minutes, dry dressed fish fillets). It indicated that sarcoplasmic protein electrophoresis patterns bands in the molecular weight region below 30 KD could be used to identify the fish species after heating 100C but not with myofibrillar protein. SDS extraction of total protein electrophoresis patterns bands also remained the same by 100C heating with the increasing time, thus it also can be used to identify the fish species but not with Urea extraction of total protein. Sarcoplasmic, myofibrillar protein and SDS, Urea extraction of total protein electrophoresis patterns bands was obviously denatured by heating at 121C with the increasing time thereby could not be used to identify the fish species. Therefore, this method does not suitable for the identification of high temperature processed fish meat products. Using the above techniques, in dry dressed fish fillet made in Lab or sold in market has shown the presence of protein band, while it did not appear in canned fish meat. This result is likely due to longer heating time and higher temperature caused the degradation of protein seriously.
This study provides the analyzing method that can effectively identify fresh filefish species by sarcoplasmic, myofibrillar protein and SDS extraction of total protein. However, this method can only be applied in the processing products of dried dressed fish fillet with slight heating processed but not in the canned fish meat products of long heating processed.

並列關鍵字

SDS-PAGE

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