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| | | | | | 第 | | 筆

詳目顯示區塊

第 25 筆
| (以作者名查詢海洋大學圖書館館藏系統)
| (以作者名查詢全國圖書書目資訊網)
| (以作者名查詢全國博碩士系統)
論文名稱: | 以固定化脂氧合酶及海藻氫過氧化物解離酶修飾海鱺肝臟油之氣味
| Odor Modification of Cobia Liver Oil Using Immobilized Lipoxygenase and Hydroperoxide Lyase from Algae
研究生: | 林安琪 An-Chi Lin
指導教授: | 孫寶年 Bonnie-Sun Pan
| 洪良邦 Liang-Bang Hung
學位類別: | 碩士
學校名稱: | 國立台灣海洋大學
系所名稱: | 食品科學系
畢業學年度: | 91
語文別: | 中文
關鍵字: | 氣味 odor
| 氫過氧化物解離酶 hydroperoxide lyase
| 脂氧合酶 lipoxygenase
全文點閱次數: | 17

全文影像連結:

電子檔1

論文頁數: | 103
摘要: | 11
| 中文摘要
| 近年箱網養殖海鱺快速增加，供食用，但其肝臟未被利用，因此
| 擬開發海鱺肝臟油，並改善其不良氣味。同一箱網養殖之海鱺，體重
| 與體長呈正相關，肥滿度、肝體比隨季節變化與魚體大小無相關性（p
| ＜0.05）。
| 以氯仿-甲醇（Folch 萃取法）、正己烷、及水煮30 分鐘萃取之肝
| 臟油，製成率分別為41.48、36.39、與28.29﹪。水煮所得肝臟油之
| 多元不飽和脂肪酸較有機溶劑萃取者高，酸價較低，魚腥味及氧化味
| 較強。
| 以固定化脂氧合酶於25℃與水煮所得之肝臟油反應2 小時，其
| 氣味可明顯改善。再配合氫過氧化物解離酶（HPLS）作用，使喜好
| 之氣味更為顯著，石蓴之HPLS 最適作用條件為pH 6.6 及35℃，於
| 37℃以下較安定，於26℃之半衰期為4 小時，於4℃為8 小時。
| 於HPLS 最適條件修飾經固定化LOX 作用後之海鱺肝油，氣味
| 之接受性均高於未處理之原油，令人喜好之氣味可能來自多元不飽和
| 脂肪酸經LOX 與HPLS 兩酵素先後作用生成之5、6、8 及9
| 個碳的醇類或醛類。香氣化合物以GC-Sniffing 及GC-MS 加
| 以鑑定， 主要之良好氣味化合物E-2-heptenal、E-2-octenal 與
| 1-octen-3-ol 含量增加， 不良氣味化合物如E,E-2,4-heptadienal、
| 2,4-heptadienal、2,6,10,14-tetramethyl pentadecane 與2,4-decadienal
| 下降。
| 以LOX 與HPLS 於25℃ 與海鱺肝臟油反應3 小時後，未
| 造成總脂肪酸含量的下降，但魚腥味明顯降低，清新之蔬果
| 甜味顯著增強，故只要少量多元不飽和脂肪酸與LOX-HPLS
| 先後作用，即能產生閾值很低的香氣化合物，改善海鱺肝臟
| 油之氣味。
|
| 13
| Abstract
| Cobia（Rachycentron canadum）has become one of the main cage
| cultured fish. In spite of the increases in processing of cobia fillet
| products, cobia liver has not been utilized commercially. Since cobia liver
| is high in lipid content, the objective of this study was to develop a cobia
| liver oil product and to improve its odor. Body weight and body length of
| cobia was significantly correlated, while condition factor, hepatosomatic
| index and seasonal changes of cobia was not correlated with body weight
| （p＜0.05）.
| The cobia liver oil was extracted with chloroform-methanol（Folch
| method）, or hexane or wet-rendered at 100℃ for 30 min. The yields
| were 41.48、36.39 and 28.29﹪respectively. The wet-rendered cobia liver
| oil was higher in polyunsaturated fatty acids, its acid value was lower and
| the odor notes of fishy and oxidized odors were stronger than those of the
| solvent extracted. The wet-rendered cobia liver oil was treated with
| oxirane-immobilized lipoxygenase （LOX） at 25℃ for 2 h, improved its
| otherwise undesirable odor.
| Treatment of this LOX-modified oil with hydroperoxide lyase
| （HPLS）, the desirable odor note increased in intensity. The optimal
| reaction condition of HPLS from a green marine macroalgae, Ulva
| conglobata, was pH 6.6 and 35℃. The algal HPLS was stable at
| temperature below 37℃. Storage at 4 and 26℃ for 24 h, the half-life was
| 8 and 4 h respectively.
| The cobia liver oil modified with LOX and HPLS at their optimal
| conditions produced desirable odorous compounds of 5、6、8 and 9
| carbons. Using GC-sniffing and GC-MS techniques, higher
| concentrations of desirable unsaturated alcohols, aldehydes, that
| resembled odors of green, melon, and fruit, were tentatively identified as
| E-2-heptenal 、E-2-octenal, 1-octen-3-ol and reduced contents of
| undesirable odor compounds including E,E-2,4-heptadienal 、
| 2,4-heptadienal、2,6,10,14-tetramethyl pentadcane and 2,4-decadienal.
| Aroma modification of cobia liver oil using LOX and HPLS at 25℃
| for 3 h, maintained 95﹪of the original polyunsaturated fatty acids along
| with a significant reduction in fishiness and higher concentration of
| fruity- and sweet- odor notes. Small amounts of polyunsaturated fatty
| acids were enough for modification of cobia liver oil odor by LOX and
| HPLS
|
論文目次: | 3
| 目錄
| 表次--------------------------------------------------------------------8
| 圖次--------------------------------------------------------------------9
| 英文代號--------------------------------------------------------------10
| 中文摘要--------------------------------------------------------------11
| 英文摘要--------------------------------------------------------------13
| 壹、前言--------------------------------------------------------------15
| 貳、文獻回顧---------------------------------------------------------17
| 一、海鱺的生態與養殖----------------------------------------------17
| 二、魚油市場---------------------------------------------------------18
| 三、魚油之萃取------------------------------------------------------19
| 四、魚油的氣味及生成途徑----------------------------------------20
| 1. 脂質自氧化作用-----------------------------------------------21
| 2. 酵素性氧化作用-----------------------------------------------21
| 五、脂氧合酶的特性---------------------------------------------------22
| 1. 自然界中之分佈-----------------------------------------------22
| 2. 作用機制--------------------------------------------------------23
| 3. 異構酶----------------------------------------------------------25
| 4
| 4. 脂氧合酶對氣味的影響----------------------------------------25
| （ a）植物氣味----------------------------------------------------25
| （ b）魚氣味-------------------------------------------------------27
| （ c）蝦氣味-------------------------------------------------------28
| （ d） 脂氧合酶作用於脂肪酸經微波加熱後形成之氣味------29
| （ e） 海藻脂氧合酶對水產品氣味的影響-----------------------29
| 六、氫過氧化物解離酶之特性-------------------------------------30
| 1. 自然界中之分佈-----------------------------------------------30
| 2. 可能之作用機制-----------------------------------------------32
| 3. 基質特異性-----------------------------------------------------33
| 4. 最適pH ---------------------------------------------------------33
| 5. 氫過氧化物解離酶產物之功能------------------------------34
| （ a）對氣味之影響-----------------------------------------------34
| （ b）與防禦系統之關係-----------------------------------------35
| 叁、材料與方法------------------------------------------------------38
| （ 一）材料------------------------------------------------------------38
| 1.海鱺肝臟---------------------------------------------------------38
| 2.海藻：石蓴------------------------------------------------------38
| （ 二）化學試劑------------------------------------------------------38
| 1.脂氧合酶----------------------------------------------------------38
| 2.固定化之載體---------------------------------------------------39
| 3.氫過氧化物------------------------------------------------------39
| 5
| （ 三）方法------------------------------------------------------------39
| 一、海鱺生理指標------------------------------------------------39
| 1.體重-----------------------------------------------------------39
| 2.體長-----------------------------------------------------------39
| 3.肥滿度--------------------------------------------------------39
| 4.肝體比--------------------------------------------------------40
| 二、肝臟油之製備------------------------------------------------40
| 1.有機溶劑萃油法---------------------------------------------40
| 2.水煮法--------------------------------------------------------40
| 3.產率-----------------------------------------------------------40
| 三、一般成分分析------------------------------------------------41
| 四、肝臟油之脂肪酸分析---------------------------------------42
| 五、酸價（AV） --------------------------------------------------43
| 六、共軛雙鍵之含量---------------------------------------------44
| 七、儲藏實驗-----------------------------------------------------44
| 八、固定化脂氧合酶的製備--------------------------------------44
| 九、固定化脂氧合酶活性之測定--------------------- ---------- 45
| 十、蛋白質定量---------------------------------------- ----------46
| 十一、氫過氧化物解離酶的萃取-------------------------------46
| 十二、氫過氧化物解離酶活性之測定－ 吸光度法------------46
| 十三、氫過氧化物解離酶特性之測定-------------------------47
| 1.最適pH--------------------------------------------------------47
| 2.最適溫度------------------------------------------------------46
| 3.熱安定性------------------------------------------------------47
| 4.儲藏安定性---------------------------------------------------47
| 6
| 5.海藻粗酵素液中HPLS 之種類與活性---------------------47
| 6.Km 及Vmax --------------------------------------------------48
| 十四、揮發性化合物之萃取、濃縮與鑑定--------------------48
| 1.揮發性化合物之萃取----------------------------------------48
| 2.揮發性成分萃取液之濃縮----------------------------------49
| 3.氣相層析（GC） ---------------------------------------------49
| 4.氣相嗅聞法（GC-Sniffing） -------------------------------50
| 5.氣相層析-質譜儀（GC-MS） 分析-------------------------51
| 十五、官能品評---------------------------------------------------51
| 1.描述性品評試驗----------------------------------------------51
| 2.經固定化LOX 作用後肝臟油之香氣----------------------51
| 3.經固定化LOX 及HPLS 作用肝臟油之香氣--------------52
| 十六、統計分析---------------------------------------------------52
| 肆、結果與討論------------------------------------------------------53
| 一、海鱺肥滿度與肝體比之季節變化--------------------------53
| 二、萃取條件對肝臟油之影響----------------------------------56
| 1.製成率---------------------------------------------------------57
| 2.肝臟油之脂肪酸組成----------------------------------------58
| 3.品質： 色澤、酸價、氣味----------------------------------64
| 4.儲藏安定性---------------------------------------------------66
| 7
| 三、石蓴氫過氧化物解離酶的特性-----------------------------68
| 1.最適pH--------------------------------------------------------70
| 2.最適溫度------------------------------------------------------72
| 3.熱安定性------------------------------------------------------73
| 4.儲藏安定性---------------------------------------------------74
| 5.種類與活性---------------------------------------------------75
| 6.Km 及Vmax --------------------------------------------------76
| 四、海鱺肝臟油經固定化LOX 及石蓴HPLS 作用後脂肪
| 酸組成之變化------------------------------------------------77
| 五、固定化黃豆LOX 及石蓴HPLS 作用時間對海鱺肝臟
| 油修飾後氣味及強度之影響-------------------------------82
| 六、揮發性化合物鑑定-------------------------------------------83
| 伍、結論--------------------------------------------------------------87
| 陸、參考文獻------------------------------------------------------ --90
| 8
| 表次
| 表一、澎湖海鱺體重、體長、肥滿度與肝體比之季節變化-------------54
| 表二、海鱺肝臟一般成分-----------------------------------------------56
| 表三、水煮法與有機溶劑法萃取海鱺肝臟油產率與回收率之比較-----57
| 表四、Folch 法與正己烷萃得海鱺肝臟油之脂肪酸組成----------------59
| 表五、以水煮法萃得海鱺肝臟油之脂肪酸組成-------------------------60
| 表六、萃取之海鱺肝臟油氣味、酸價、顏色及強度之比較-------------65
| 表七、石蓴（Ulva conglobata）氫過氧化物解離酶的活性---------75
| 表八、海鱺肝臟油經固定化黃豆脂氧合酶及氫過氧化物解離酶作用後
| 不同飽和度之脂肪酸含量百分比之變化-------------------------77
| 表九、經固定化黃豆脂氧合酶及氫過氧化物解離酶作用海鱺肝臟油後
| 脂肪酸組成之變化-----------------------------------------------80
| 表十、固定化黃豆脂氧合酶及石蓴（Ulva conglobata）氫過氧化物解
| 離酶作用時間對海鱺肝臟油修飾氣味及強度之影響-------------81
| 表十一、利用減壓冷凝法收集及氣相層析-質譜儀和氣相層析嗅聞法
| 鑑定修飾前後海鱺肝臟油之良好氣味成份-----------------------84
| 表十二、利用減壓冷凝法收集及氣相層析-質譜儀和氣相層析嗅聞法
| 鑑定修飾前後海鱺肝臟油之不良氣味成份-----------------------86
| 附表一、民國82-90 年台灣海鱺生產量及肝臟重量--------------------89
| 9
| 圖次
| 圖一、海鱺體重與體長之關係------------------------------------------54
| 圖二、海鱺體重與肥滿度之關係----------------------------------------55
| 圖三、海鱺體重與肝體比之關係----------------------------------------55
| 圖四、水煮法與有機溶劑萃取之海鱺肝臟油脂肪酸組成之差異--------63
| 圖五、海鱺肝臟油於-20 ℃儲藏四個月中氧化指標A 234 nm 之變化----67
| 圖六、海鱺肝臟油於-20 ℃儲藏四個月中氧化指標A 268 nm 之變化----67
| 圖七、石蓴（Ulva conglobata）氫過氧化物解離酶活性隨作用時間
| 之變化-----------------------------------------------------------68
| 圖八、石蓴（Ulva conglobata）粗酵素液之蛋白質量與氫過氧化物
| 解離酶活性之關係---------------------------------------------69
| 圖九、石蓴（Ulva conglobata）氫過氧化物解離酶之最適pH ---------71
| 圖十、石蓴（Ulva conglobata）氫過氧化物解離酶之最適溫度--------72
| 圖十一、石蓴（Ulva conglobata）氫過氧化物解離酶之熱安定性-----73
| 圖十二、石蓴（Ulva conglobata）氫過氧化物解離酶於4 及26℃
| 下之儲藏安定性--------------------------------------------74
| 圖十三、石蓴（Ulva conglobata）氫過氧化物解離酶與經固定化
| 脂氧合酶作用之海鱺肝臟油於pH 6.6、26℃ 反應4
| 分鐘之Lineweaver-Burk 圖-------------------------------76
| 18:2：linoleic acid
| 18:3：linolenic acid
| 20:4：arachidonic acid
| ADH：alcohol dehydrogenase
| DHA：docosahexaenoic acid
| EPA：eicosapentaenoic acid
| HODE：hydroxyoctadecadienoic acid
| HPETE：hydroperoxy-eicosatetraenoic acid
| HPLS：hydroperoxide lyase
| HPO：hydroperoxides
| HpODE：hydroperoxyoctadecadienoic acid
| HPOT：hydroperoxyoctadecatrienoic acid
| HUFA：highly unsaturated fatty acid
| LOX：lipoxygenase
| PUFA：polyunsaturated fatty acid
|
參考文獻: | 90
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