臺灣博碩士論文加值系統

本研究以取自母牛分娩後第二至四天分泌且等體積混合的乳汁作為材料，以阿拉伯膠、β-環狀糊精和幾丁聚醣之10 %水溶液，依1:4體積比和牛初乳或乳清混合，以冷凍及噴霧乾燥法進行微膠囊化，並探討pH值、溫度、胃腸道蛋白酶、微生物脂多醣及儲藏時間對微膠囊化初乳與乳清IgG活性之影響。
結果顯示冷凍乾燥法所得之微膠囊化初乳其IgG活性比噴霧乾燥法所得者高。在酸鹼安定性方面，未微膠囊化之初乳在pH 7~8有較高之IgG活性，經微膠囊化者則以阿拉伯膠所得者在pH 7~8時有較高之IgG活性。在熱安定性方面，阿拉伯膠和β-環狀糊精對於初乳或乳清之IgG均具有保護作用。在胃腸道蛋白酶耐受性方面，以阿拉伯膠及β-環狀糊精微膠囊化之初乳經胃蛋白酶作用2小時後，其IgG殘存活性較未微膠囊化者分別高出9.8及7.6 %，再經胰蛋白酶作用4小時後，有、無微膠囊化處理則無顯著之差異性；而微膠囊化對乳清之胃蛋白酶及胰蛋白酶耐受性則無顯著影響。在與微生物脂多醣作用方面，E. coli O55:B5與初乳或乳清之IgG有抗原-抗體親和性反應，而以阿拉伯膠微膠囊化者，在經過胃蛋白酶作用後，其與E. coli O55:B5脂多醣作用後之IgG活性較未微膠囊化者高。在儲藏安定性方面，儲藏溫度與包裝材質對於初乳或乳清之IgG殘存活性有顯著影響，在4 ℃儲存60天後之IgG活性較室溫高，以鋁袋包裝儲存者高於以透明塑膠袋包裝者，以阿拉伯膠微膠囊化之初乳在4 ℃儲存60天後之IgG殘存活性顯著高於未微膠囊化者。

The bovine colostrums collected from the second to the fourth day postpartum were mixed by equal volume and used as raw materials. A 10 % aqueous solution of gum arabic, β-cyclodextrin (β-CD) or
chitosan was added to an equal volume of colostrums or whey, and microencapsulated through freeze drying and spray dryng methods. The effects of pH value, temperature, pepsin and trypsin, lipopolysaccharides (LPS) from E. coli (serotype O55:B5), and storage time on the IgG activity of microencapsulated colostrums and whey were investigated.
Results showed that microencapsulated colostrums prepared through freeze drying had higher IgG activity than those prepared through the spray drying. The results for the effects of the five factors on the IgG activity of microencapsulated colostrums and whey were summarized as follows: (1) pH stability: The non-microencapsulated colostrums were found to have higher IgG activity at a pH value of 7~8 than the microencapsulated colostrums; the microencapsulated colostrums with gum arabic were found to have higher IgG residual activity at a pH value of 7~8. (2) Thermal stability: Gum arabic and β-cyclodextrin (β-CD) were found to have a protective effect on the IgG activity of colostrums and whey. (3) Pepsin and trypsin Tolerance: Under the action of pepsin for two hours, the residual IgG activities of microencapsulated colostrums with gum arabic and β-cyclodextrin (β-CD) were higher by 9.8 % and 7.6 %, respectively, than that of non-microencapsulated colostrums. After further action by trypsin for four hours, no significant differences were found between microencapsulated and non-microencapsulated colostrums. However, the action of pepsin and trypsin did not significantly affect the IgG residual activity of microencapsulated whey. (4) Action of LPS from microorganisms: The E. coli (serotype O55:B5 form) was found to have an antigen-antibody affinity with the IgG of colostrums and whey. However, after the action of pepsin and the effect of LPS from E. coli (serotype O55:B5 form), the microencapsulated colostrums with gum arabic was found to have higher IgG activity than the non-microencapsulated colostrums. (5) Storage stability: Storage temperature and packaging materials were found to have a significant impact on the residual IgG activity of colostrums and whey. After being stored for 60 days at 4 ℃, the IgG activity was found to be higher than that stored at room temperature. As for packaging materials, aluminum bag-packaged colostrums and whey were found to have a higher IgG activity than transparent plastic bag-packaged colostrums. After storage for 60 days at 4 ℃, the microencapsulated colostrums with gum arabic were found to have a significantly higher IgG residual activity than the non-microencapsulated colostrums.

封面內頁
簽名頁
授權書iii
中文摘要iv
英文摘要vi
誌謝viii
目錄ix
圖目錄xii
表目錄xv
第一章緖言1
第二章文獻回顧3
2.1牛初乳3
2.1.1牛初乳簡介3
2.1.2牛初乳的成分4
2.1.3牛初乳中蛋白質組成4
2.2免疫系統7
2.2.1免疫系統及其反應7
2.2.2抗體簡介9
2.2.3抗體的種類10
2.2.4抗體之防禦機制11
2.2.5免疫球蛋白之安定性15
2.3免疫球蛋白G (IgG)之分析法16
2.3.1定量分析16
2.3.2定性分析19
2.4微膠囊化19
2.4.1微膠囊化定義19
2.4.2微膠囊化方法20
2.4.3微膠囊化包覆粉體種類24
第三章材料與方法32
3.1實驗材料32
3.1.1樣品製備32
3.1.2包覆粉體32
3.1.3試藥33
3.1.4儀器33
3.2實驗方法34
3.2.1基本組成分析34
3.2.2牛初乳與乳清之微膠囊化38
3.2.3電子顯微鏡之觀察41
3.2.4微膠囊化IgG安定性之測定41
3.2.5IgG對胃腸蛋白酶之耐受性42
3.2.6牛初乳和乳清與微生物脂多醣之親和性43
3.2.7統計分析44
第四章結果與討論45
4.1初乳和乳清之一般基本組成45
4.2初乳IgG之定量分析47
4.3微膠囊化處理47
4.3.1不同微膠囊化方法之比較47
4.3.2電子顯微鏡觀察結果50
4.4酸鹼對IgG安定性影響53
4.5熱處理對IgG安定性影響58
4.6胃腸消化酵素對IgG之影響62
4.7與微生物脂多醣作用之親和力64
4.8初乳與乳清IgG之儲藏安定性68
第五章結論76
參考文獻78
圖目錄
圖2.1免疫球蛋白的結構：(a)分子結構(b)分子褶疊的特徵結構13
圖2.2微膠囊的各種包覆形式22
圖2.3α、β及γ-環狀糊精結構27
圖2.4幾丁質及幾丁聚醣之化學結構30
圖3.1本研究之實驗流程35
圖3.2酵素連接免疫吸附分析法39
圖4.1牛血清IgG以ELISA法所得之標準曲線48
圖4.2牛血清IgG以ELISA法所得之標準曲線及線性回歸之R-square(0.9882)48
圖4.3以不同乾燥法進行微膠囊化所得初乳之IgG殘存活性49
圖4.4以不同乾燥法進行微膠囊化所得初乳之IgG殘存活性51
圖4.5初乳(A)及以阿拉伯膠(B)、β-環狀糊精(C)、幾丁聚醣(D)微膠囊化之初乳掃描式電子顯微照片52
圖4.6乳清(A)及以阿拉伯膠(B)、β-環狀糊精(C)、幾丁聚醣(D)微膠囊化之乳清掃描式電子顯微照片54
圖4.7在不同pH值下之初乳和微膠囊化初乳之IgG活性55
圖4.8在不同pH值下之乳清和微膠囊化乳清之IgG活性56
圖4.9在pH 7和8下之初乳及微膠囊化初乳之IgG殘存活性57
圖4.10在pH 7和8下之乳清及微膠囊化乳清之IgG殘存活性59
圖4.11初乳及微膠囊化初乳分別經63.5 ℃加熱30 min及75 ℃加熱15 sec後之IgG殘存活性60
圖4.12乳清及微膠囊化乳清分別經63.5 ℃加熱30 min及75 ℃加熱15 sec後之IgG殘存活性61
圖4.13初乳和乳清於不同pH值下經pepsin作用1、2、4、8小時後之IgG殘存活性63
圖4.14初乳及微膠囊化初乳以胃蛋白酶作用2小時及胰蛋白酶作用4小時後之IgG殘存活性65
圖4.15乳清及微膠囊化乳清以胃蛋白酶作用2小時及胰蛋白酶作用4小時後之IgG殘存活性66
圖4.16初乳與乳清經E. coli O55:B5脂多醣及兔子抗牛血清IgG反應後之ELISA值67
圖4.17未微膠囊化及以阿拉伯膠微膠囊化之初乳與乳清經胃蛋白酶和胰蛋白酶水解後，再經E. coli O55:B5脂多醣作用後之IgG殘存活性69
圖4.18未微膠囊化及以阿拉伯膠微膠囊化之初乳和乳清以透光塑膠袋包裝並在不同溫度(4 ℃和室溫)下儲存60天期間之IgG殘存活性70
圖4.19未微膠囊化及以阿拉伯膠微膠囊化之初乳和乳清以鋁袋包裝並在不同溫度(4 ℃和室溫)下儲存60天期間之IgG殘存活性72
圖4.20未微膠囊化與微膠囊化初乳和乳清在室溫下以透光塑膠袋和鋁袋儲存60天後之IgG殘存活性73
圖4.21未微膠囊化與微膠囊化初乳和乳清在4 ℃和室溫下以透光塑膠袋儲存60天後之IgG殘存活性74
表目錄
表2.1初乳與常乳的成分5
表2.2微膠囊包覆各種技術23
表4.1初乳和乳清一般基本組成46

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