臺灣博碩士論文加值系統

魚油中的魚腥味主要來自多元不飽和脂肪酸 ( PUFA ) 之氧化，為了增進魚油的應用性，本研究針對市售食用魚油進行脂肪酸組成與氣味品質分析，再以固定化海藻脂氧合酶修飾魚油氣味，期增加魚油之應用性。

魚油以脂肪酸甲酯化法中反應條件較激烈的BF3 ( boron trifluoride ) 及反應條件較緩和的TMAH ( tetramethylammonium hydroxide ) 酯化後，分別進行氣相層析。由於BF3法以熱迴流進行轉酯化，可將魚油中TG及乙酯化態的EPA與DHA轉為甲酯化態，而TMAH法在室溫反應，僅能甲酯化TG型態的EPA與DHA，經層析圖比較後，顯示美國產魚油樣品膠囊中含TG及乙酯化態之EPA與DHA兩種型態。

以BF3酯化法分析，市售食用魚油膠囊之EPA與DHA總合佔其脂肪酸總量之15-75 %。市售魚油膠囊內之魚油具魚腥味，氣味強度差異甚大。另添加保健成分之複方魚油，其添加物有銀杏、大蒜萃取物、紅麴萃取物、植物醇、卵磷脂、γ-胺基丁酸等，其魚油的腥氣較單方魚油強。

石蓴脂氧合酶修飾魚油氣味，自2004年8月至2005年4月葉狀體製成粗酵素液，其脂氧合酶比活性範圍在1.14 - 2.90 μmol/mg-min。颱風後採集之石蓴，LOX活性高達4.95 μmol/mg-min。採集退潮後水面下之石蓴，其活性範圍介於1 - 2.90 μmol/mg-min。石蓴脂氧合酶以硫酸銨部分純化，在飽和濃度40 - 60 % 的劃分範圍具有活性，在透析液中添加0.2 mM Ca2+可以提升酵素的活性，回收率為59.72 %。

石蓴粗酵素液及經過硫酸銨飽和濃度40-60 % 部分純化之石蓴LOX，分別與載體oxirane acrylic beads混合，於室溫中75 rpm震盪20小時進行固定化處理。固定化之石蓴LOX與0.5μmole亞麻油酸作用於25℃、以75 rpm 震盪反應，反應需2小時達平衡，酵素活性高於固定化之石蓴粗酵素液。固定化之石蓴LOX與市售膠囊魚油於25℃、75 rpm震盪反應2小時後，發現有好的氣味產生，這些氣味分子的化學結構以GC/MS鑑定，為甜瓜味 ( Ethyl acetate )、水果味 ( E,E-3,5-Octadiene-2-one ) 等。

表目錄 x
圖目錄 xii
貳、文獻回顧 3
一、魚油的特性 3
1.脂肪酸組成 3
2.安定性 3
3.營養價值 4
二、油脂氣味的形成 5
1.脂肪酸的氧化 5
2.氧化產物 6
3.揮發性成分 7
三、氣味分析方法 8
四、脂氧合酶與水產品氣味的關係 14
1.酵素與魚油作用後的氣味 14
2.修飾氣味的應用 14
五、脂氧合酶固定化方式與特性 15
1.固定化方式 15
2.固定化LOX的特性 17
參、材料與方法 19
一、材料 19
1.市售魚油 19
2.海藻 19
3.Oxirane acrylic beads 19
二、分析方法 19
1.石蓴脂氧合酶酵素液之製備 19
(1)緩衝溶液 19
(2)石蓴脂氧合酶粗酵素液萃取 20
(3)硫酸銨分劃 20
2.脂氧合酶活性測定 - 吸光度法 ( LOOH 產物測定 ) 21
3.蛋白質定量 - 染料結合法 ( dye binding ) 22
4.固定化酵素的製備 - 化學共價鍵法 22
5.固定化脂氧合酶活性之測定 23
6.固定化酵素反應時間測定 23
7.魚油修飾後之香氣品評 23
8. 揮發性化合物鑑定- Headspase Solid Phase Microextraction法 24
(1)氣味化合物之萃取 24
(3)氣液相層析-質譜儀GC/MS分析 25
(4)滯留指數 ( retention index, R.I. ) 測定 25
9.脂肪酸分析 27
(1)甲基酯化 27
(2)氣相層析分析 28
肆、結果與討論 29
一、市售魚油脂肪酸之組成 29
1. 魚油產品的分類 29
2. 魚油脂肪酸之組成 39
二、石蓴脂氧合酶之固定化 44
1. 石蓴脂氧合酶之活性 44
2. 脂氧合酶之部分純化 47
3. 固定化石蓴脂氧合酶 51
三、石蓴脂氧合酶對魚油氣味之修飾 53
1. 石蓴香氣化合物之鑑定 53
2. 魚油氣味化合物之鑑定 56
3. 修飾魚油之脂肪酸組成與氣味鑑定 62
陸、結論 68
柒、參考文獻 70

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