臺灣博碩士論文加值系統

第一部分
本研究目的為探討以益生菌發酵大豆粕，依不同比例來取代魚粉做為飼料。以不同取代配方飼料分別餵養吳郭魚和鱸魚，探討對其成長之影響。本研究之控制組飼料以市售魚粉為飼料中蛋白質來源；而實驗組分為兩部分，第一是六組分別是以未發酵大豆粕取代20%及40%魚粉、A菌發酵大豆粕及P菌發酵大豆粕之來飼育吳郭魚，分別簡稱為SBM20、SBM40、FLA20、FLA40、FLP20與FLP40；另一是以A菌發酵大豆粕取代15%、30%及45%之魚粉來飼育金目鱸，分別簡稱為FLA15、FLA30及FLA45，進行六十天的飼育試驗。吳郭魚和金目鱸飼育試驗結果以存活、成長狀況(增重、增重百分比、比生長率)、飼料消化利用(飼料轉換率、飼料效率)及體內組成。由結果顯示吳郭魚在增重及內臟身體比皆以 SBM20 最佳，成長狀況以控制組飼料為最佳，而飼料消化利用以FLP20最佳。而金目鱸在增重、內臟身體比及肝體比是以控制組飼料最佳，而成長狀況則以 FLA15 為最佳，飼料消化利用則以 FLA30 最佳。另外，胺基酸組成中之必需胺基酸含量皆為 FLA30 為最高。

第二部分
本實驗目的是將魚卵經酵素水解後，進行脫脂製成魚卵水解物 (roe hydrolyzate, RH)，然後進行純化。首先，以陰離子交換管柱進行區分，再以膠體過濾層析(G-25)進行分離純化，結果以A區區分物之抑制酪胺酸酶活性最佳，達到78.20 % 抑制效果。最後以高效液相層析為C18管柱將A區區分物進一步的分離，進行純化抑制酪胺酸酶活性胜肽之胺基酸定序。

PartⅠ.
The purpose of this study was to explore the replacement of fish meal with the probiotic fermented soybean meals as feed and to investigate the effects of different feeds on the growth of tilapia and giant seaperch (Lates calcarifer). There were two animal experiments. The tilapia feeds, comprising widely available and affordable raw ingredients, were iso-energetic and iso-carbohydrated, but contained protein of different sources as follows: 100% of protein from fishmeal, protein from fishmeal (80 or 60%) plus from soybean meal (20 or 40%), protein from fishmeal (80 or 60%) plus from bacterium A fermented soybean meal (20 or40%), and protein from fishmeal (80 or 60%) plus from bacterium P fermented soybean meal (20 or40%). They were called control, FLA20, FLA40, FLP20, FLP40, SBM20, and SBM40 groups, respectively. The giant seaperch feeds contained protein of different sources as follows: 100% of protein from fishmeal (control), 85% of protein from fishmeal and 15% of protein from bacterium A fermented soybean meal (FLA15), 70% of protein from fishmeal and 30% of protein from bacterium A fermented soybean meal (FLA30), and 55% of protein from fishmeal and 45% of protein from bacterium A fermented soybean meal (FLA45). Both two experiments were conducted 60-day feed trials. The survival rate, growth performance (weight gain, percentages weight gain, specific growth rate), feed digestion (feed conversion rate, feed efficiency) and body composition of two fish groups were investigated. The results indicate that SBM20 group exhibited the highest weight gain and visceral somatic index among tilapia groups. Control diet demonstrated the best growth performance. FLP20 demonstrated the best feed digestion and utilization with the experimental diets. In the seaperch experiment, the control group exhibited the highest weight gain, visceral somatic index and hepatosomatic index among seaperch group. FLA15 and FLA30 group exhibited the best growth performance and the best feed digestion and utilization among all group, respectively. In addition, the amount of essential amino acids in amino acid composition of FLA30 is the highest among all groups.

PartⅡ.
In this experiment, roe hydrolysate (RH) was produced from roes treated with enzymatic hydrolysis and defatting. To screen peptides with tyrosinase inhibitory activity, RH was fractionated and purified by anion exchange chromatography and then gel filtration (G-25) chromatography. A fraction with 78.20% tyrosinase inhibitory activity was obtained. Finally C18 HPLC column was used for further separation. A tyrosinase inhibitory peptide was purified and its amino acid sequence. Tyrosinase inhibitory peptides can be used in the development of skin care products.

摘要-第一部分 I
摘要-第二部分 II
Abstract III
PartⅠ. III
PartⅡ. V
誌謝 VI
目錄 VIII
表目錄 XV
圖目錄 XVI
附目錄 XIX
符號說明 XX
壹、前言 1
第一部分、發酵大豆取代部分魚粉之飼料配方開發 1
第二部分、魚卵水解物抑制酪胺酸酶活性探討 3
貳、文獻整理 4
第一部分、發酵大豆取代部分魚粉之飼料配方開發 4
一、吳郭魚簡介 4
(一) 吳郭魚之分類 5
(二) 吳郭魚之特性 7
(三) 吳郭魚之養殖概況 7
二、金目鱸簡介 8
(一) 金目鱸之分類 8
(二) 金目鱸之特性與生態 9
(三) 金目鱸之養殖概況 10
三、魚粉 (Fish meal) 的近況 11
四、大豆粕 (Soy Bean Meal, SBM) 簡介 12
五、益生菌 (Probiotics) 簡介 18
六、Lactobacillus乳酸菌屬 20
第二部分、魚卵水解物抑制酪胺酸酶活性探討 21
一、黑色素 (Melanin) 之形成 21
参、材料與方法 23
第一部分、發酵大豆取代部分魚粉之飼料配方開發 23
一、實驗材料 24
1. 材料來源 24
2. 實驗藥品 25
3. 儀器設備 28
二、實驗方法 30
1. 大豆粕(Soybean meal, SBM)前處理 30
1.1 蒸煮大豆粕製備 30
1.2 未發酵大豆粕(SBM)之製備 30
1.3 發酵大豆粕(Fermented soybean meal, FSBM)預製備 30
1.3.1 MRS broth 培養液製備 30
1.3.2 發酵工作液製備 32
1.4 發酵大豆粕 (FSBM) 製備 32
2. 一般成分分析 34
2.1 水分 (Moisture) 34
2.2 灰分 (Ash) 35
2.3 粗蛋白 (Crude protein) 35
2.4 粗脂肪 ( Crude fat ) 36
3. 吳郭魚飼育試驗 38
3.1 實驗用魚 38
3.2 實驗方法 38
3.2.1 飼育動物及環境條件 38
3.2.2 吳郭魚飼料配方及製備 39
3.3 魚體資料分析 42
3.4 魚體組織分析 43
3.4.1 一般成分分析 43
4. 金目鱸飼育試驗 45
4.1 實驗用魚 45
4.2 實驗方法 45
4.2.1 飼育動物與環境條件 45
4.2.2 金目鱸飼料配方及製備 46
4.3 魚體資料分析 48
4.4 魚體組織分析 48
第二部分、魚卵水解物抑制酪胺酸酶活性探討 50
1. 魚卵 (Roe) 前處理 51
2. 魚卵水解物 (Roe hydrolyate, RH) 51
3. RH純化 51
3.1 快速蛋白質液相層析 52
3.1.1 陰離子交換層析 52
3.1.2 膠體過濾層析 53
3.2 高效液相層析 (High performance liquid chromatography, HPLC) 54
3.2.1 C18 半製備型和分析型管柱 54
3.3 胺基酸定序 55
4. 抗酪胺酸酶活性測定 56
5. 統計分析 57
肆、結果與討論 58
第一部分、發酵大豆取代部分魚粉之飼料配方開發 58
一、 實驗飼料一般成分分析 58
二、吳郭魚飼育實驗之成長表現 61
(一) 吳郭魚餵食實驗飼料60天後之存活率(%) 61
(二)吳郭魚餵食實驗飼料60天後之增重 63
(三)吳郭魚餵食實驗飼料60天後之增重百分比 (PWG) 65
(四)吳郭魚餵食實驗飼料60天後之飼料轉換率(FCR) 67
(五)吳郭魚餵食實驗飼料60天後之飼料效率 (FE) 69
(六)吳郭魚餵食實驗飼料60天後之比生長率 (SGR) 71
(七)吳郭魚餵食實驗飼料60天後之肌肉率(MR) 73
(八)吳郭魚餵食實驗飼料60天後之內臟比 (VSI) 75
(九)吳郭魚餵食實驗飼料60天後之肝體比 (HSI) 77
(十)吳郭魚餵食實驗飼料60天後之魚體組織成分 79
(十一)吳郭魚餵食實驗飼料60天後之魚體胺基酸分析 81
三、金目鱸飼育實驗之成長表現 83
(一) 金目鱸餵食實驗飼料 60天後之存活率 83
(二) 金目鱸餵食實驗飼料 60天後之增重 (WG) 85
(三) 金目鱸餵食實驗飼料 60天後之增重百分比 (PWG) 87
(四) 金目鱸餵食實驗飼料 60天後之飼料轉換率(FCR) 89
(五) 金目鱸餵食實驗飼料 60天後之飼料效率(FE) 91
(六) 金目鱸餵食實驗飼料 60天後之比生長率 (SGR) 93
(七) 金目鱸餵食實驗飼料 60天後之肌肉率 (MR) 95
(八) 金目鱸餵食實驗飼料 60天後之內臟身體比 (VSI) 97
(九) 金目鱸餵食實驗飼料 60天後之肝體比 (HSI) 99
(十) 金目鱸餵食實驗飼料 60天後之魚體組織分析 101
(十一) 金目鱸餵食實驗飼料 60天後之魚體胺基酸分析 103
第二部分、魚卵水解物抑制酪胺酸酶活性探討 105
一、 RH 陰離子交換層析及膠體過濾層析區分物純化之探討 105
二、 純化 RH 區分物之抗酪胺酸酶活性探討 109
(一) RHⅡA 之抗酪胺酸酶活性探討 114
伍、結論 117
第一部分、發酵大豆取代部分魚粉之飼料配方開發 117
第二部分、魚卵水解物抑制酪胺酸酶活性探討 118
陸、參考文獻 119
中文文獻 119
英文文獻 123
柒、附錄 130

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