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論文名稱: | 藍點鸚哥魚粉紅色腸道色素之鑑定與體內藻蛋白形式之變化
| Identification of pigment in pink gut and changes of phycobiliprotein forms in parrotfish (Scarus ghobban).
研究生: | 黃慧茹 Hui-Ju Huang
指導教授: | 孫寶年 Bonnie Sun Pan
| 陳麗淑
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
學號: | M91320019
畢業學年度: | 92
語文別: | 中文
關鍵字: | 藍點鸚哥魚 parrotfish
| 藻紅蛋白 phycoerythrin
| 藻藍蛋白 phycocyanin
| 異藻藍蛋白 allophycocyanin
| 螢光 fluorescence
| 粉紅色腸道 pink gut
全文點閱次數: | 3

全文影像連結:

論文全文(pdf)

論文頁數: | 61
摘要: | 摘    要
|
| 鸚哥魚（Parrotfish, Scaridae）是珊瑚礁區優勢的草食性魚類。剛捕獲之鸚哥魚的腸道後段經常出現粉紅色，經蓄養一段時間後粉紅色會褪去。經分光光度計分析腸道粗萃取液之吸收光譜，及以共軛焦點顯微鏡觀察其組織切片之螢光分佈，推測此粉紅色物質的來源為紅藻。本研究將進一步鑑定此呈色物質，並探討鸚哥魚對紅藻的消化後，紅藻的代謝及變化。
|
| 測定藍點鸚哥魚（Scarus ghobban）成魚的粉紅色腸道、血漿及魚皮中色素的吸收光譜，並與紅藻萃取液比對。粉紅色腸道萃取液的最大吸收波峰在490及565 nm，與紅藻萃取液的最大吸收波峰相同，而血漿的吸收波長在424、540、575和650 nm，魚皮萃取液的最大吸收波峰則是在650 nm，其中血漿和魚皮萃取液在650 nm有相同的吸收波峰。
|
| 進一步比較其螢光物質，紅藻萃取液與粉紅色腸道萃取液在575 nm有相同的發射波峰，而血漿和魚皮萃取液在650 nm有相同的發射波峰，此二發射波峰分別與藻紅蛋白的發射波峰575 nm及藻藍蛋白之發射波峰650 nm相同，因此推測粉紅色腸道、血漿及魚皮中之色素分別由藻紅蛋白及藻藍蛋白等藻膽蛋白構成。
| 紅藻萃取液和粉紅色腸道萃取液的紅色素分子量相同，約為227 kDa，而血漿及魚皮中的色素分子量為125 kDa，鸚哥魚在食入紅藻後，藻紅蛋白在中腸後段堆積，經消化吸收，進入血液後轉為分子量較小的藻藍蛋白，運送至魚皮呈現藍色。
|
| Abstract
|
| Parrotfish (Parrotfish, Scaridae) is a dominant species of herbivorous fishes living in coral reefs. The posterior intestine of parrotfish is usually pink when the parrotfish is caught alive and sacrifice instanly. The pink color disappears after the fish are cultivated in aquaria for a few days. The absorption spectra of the pink gut extract and the confocal microscopic observation of the gut tissue, the ingested red algae seems to be the source of pink gut. The objective of this study is to identify the pigment and its possible metabolic changes during ingestion and digestion of the red algae.
|
| The pink gut of the blue-barred parrotfish (Scarus ghobban)and red algae (Gelidium pacificum) were extracted with phosphate buffer respectively and showed similar absorption spectra. The λmax was 490, 565 nm similar to those of phycoerythrin. The extracts of plasma, skin and allophycocyanin had same λmax at 650 nm. The fluorescent spectra of the extracts of red algae, pink gut and phycoerythrin all had λem at 575 nm. In addition to a λem at 650 nm for plasma, skin allophycocyanin.
|
| The molecular weight of the pigments determined by native- polyacrylamide gel electrophoresis, was 227 kDa for that of the red algae and the pink gut extract. Those of the plasma and the skin extract had a molecular weight of 125 kDa. It is thus supposed that the blue-barred parrotfish eat red algae and the phycoerythrin is storaged at posterior intestine. After digestion, the phycoerythrin turned into phycocyanin or allophycocyanin and get transported by blood, appears blue to skin, which in color.
|
論文目次: | 目    錄
| 英文摘要    Ⅰ
| 中文摘要    Ⅱ
|
| 壹、前言    1
| 貳、文獻回顧    2
| 一、鸚哥魚的生理及生態    2
| （一）生活史    2
| （二）消化道型態及特徵    3
| （三）攝食    3
| 二、海藻的分類及分佈    4
| （一）海藻    4
| （二）草皮狀海藻    5
| 三、藻膽蛋白之種類及特性    6
| （一）藻紅蛋白    6
| （二）藻藍蛋白與異藻藍蛋白    9
| 參、材料與方法    11
| 一、材料    11
| （一）鸚哥魚    11
| （1）幼魚    11
| （2）成魚    11
| （二）紅藻    14
| 二、方法    16
| （一）色素之萃取    16
| （二）體表色澤之測定    17
| （三）吸收光譜及螢光光譜之測定    17
| （四）分子量之測定－Native - PAGE    18
| （五）養殖實驗－餵食紅藻及綠藻    18
| （六）一般成份分析    20
| （七）蛋白質定量    23
| （八）統計分析    23
| 肆、結果與討論    24
| ㄧ、魚體色素與藻蛋白吸收光譜之比較    24
| （一）紅藻與粉紅色腸道－藻紅蛋白    24
| （二）血漿與魚皮－藻藍蛋白    26
| 二、魚體色素之螢光    29
| （一）紅藻與粉紅色腸道    29
| （二）血漿與魚皮    34
| 三、藍點鸚哥魚體內藻蛋白形式之轉變    42
| 四、攝食紅藻或綠藻對體表色澤與色素之影響    46
| 伍、結論    50
| 陸、參考文獻    51
|
| 圖    次
| 圖一、藻紅蛋白呈色基團phycoerythrobilin（PEB）與phycourobilin（PUB）之化學結構。(a) single-linked PEB; (b) double-linked PEB; (c) double-linked PUB -----------------------------------------8
| 圖二、藻藍蛋白呈色基團phycocyanobilin（PCB）與phycobiliviolin（PXB）之化學結構。(a) PCB; (b) PXB ------------------------10
| 圖三、藍點鸚哥魚（Scarus ghobban）初期型成魚（IP）之外觀
| 12
|
| 圖四、藍點鸚哥魚之腸道分段，食道 (Ⅰ)、小腸囊組織 (Ⅱ)、前腸(Ⅲ)、中腸前段(Ⅳ)、中腸後段(Ⅴ)及後腸(Ⅵ)，第Ⅴ段腸道呈現粉紅色----------------------------------------------------------------12
| 圖五、太平洋石花菜（Gelidium pacificum）------------------------------15
| 圖六、太平洋石花菜（Gelidium pacificum）萃取液（pH 7.0之 0.05 M磷酸鹽緩衝液）之吸收光譜 ---------------------------------------24
| 圖七、粉紅色腸道萃取液（pH 7.0之 0.05 M磷酸鹽緩衝液）之吸收光譜    25
| 圖八、藍點鸚哥魚血漿呈色（血液於4℃經2000 × g離心10 min之上層液）（A）無粉紅色腸道（B）有粉紅色腸道    26
| 圖九、藍點鸚哥魚血漿之吸收光譜（血液於4℃經2000 × g離心10 min，取上層液）    27
| 圖十、藍點鸚哥魚之魚皮萃取液之吸收光譜    28
| 圖十一、太平洋石花菜萃取液以374 nm為激發波長之發射光譜    30
| 圖十二、太平洋石花菜萃取液以495 nm為激發波長之發射光譜    31
| 圖十三、粉紅色腸道萃取液以377 nm為激發波長之發射光譜    32
| 圖十四、粉紅色腸道萃取液以495 nm為激發波長之發射光譜    33
| 圖十五、藍點鸚哥魚血漿以319 nm為激發波長之發射光譜    35
| 圖十六、有粉紅色腸道之藍點鸚哥魚血漿以349 nm為激發波長之發射光譜    36
| 圖十七、魚皮萃取液以285 nm為激發波長之發射光譜    37
| 圖十八、魚皮萃取液以322 nm激發之發射光譜    38
| 圖十九、藍點鸚哥魚魚體各部位色素與紅藻萃取液之10％ Native-PAGE    42
| 圖二十、藍點鸚哥魚魚體各部位色素與紅藻萃取液之10％ Native-PAGE(經染色)    44
| 圖二十一、Native-PAGE電泳Marker 分子量與Rf值之對數比對圖    45
| 圖二十二、藍點鸚哥魚幼魚及成魚之外觀與體色    46
|
| 表    次
| 表一、添加紅藻之鸚哥魚飼料配方    19
| 表二、藍點鸚哥魚含紅藻飼料之一般成分    22
| 表三、藍點鸚哥魚粉紅色腸道與紅藻萃取液(1)激發波長（λ ex）與發射波峰（λ em）之比較    34
| 表四、藍點鸚哥魚魚皮萃取液(1)之激發波長（λ ex）及發射波峰（λ em）    39
| 表五、藍點鸚哥魚血漿色素之激發波長（λ ex）及發射波峰（λ em）    40
| 表六、藍點鸚哥魚幼魚之生物指標    48
| 表七、藍點鸚哥魚幼魚及成魚體表色澤之差異    49
|
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