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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 蔣丙煌(Been-Huang Chiang) | |
| dc.contributor.author | Jia-Ying Lin | en |
| dc.contributor.author | 林佳瑩 | zh_TW |
| dc.date.accessioned | 2021-06-16T05:11:22Z | - |
| dc.date.available | 2019-08-21 | |
| dc.date.copyright | 2014-08-21 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-19 | |
| dc.identifier.citation | 詹玲、潘思軼。大豆糖蛋白的分離純化及結構分析。食品科學。2007，27，594-596。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/55945 | - |
| dc.description.abstract | 自然殺手細胞為負責對抗病毒感染及腫瘤細胞的第一道防線，在免疫系統中扮演著關鍵的角色。近年來以飲食調節免疫功能為研究的一大趨勢，由天然來源分離出的醣蛋白被證實具有免疫調節的潛力。黃豆富含高量蛋白質且乳酸菌發酵黃豆產品已被證實具有免疫調節活性。然而，目前之發酵產品多以液態發酵方式生產為主。本研究則以黃豆粉為基質，以兩階段式固態發酵方式，先以 Saccharomyces cerevisiae 預發酵後，再接種 Lactobacillus acidophilus、Streptococcus thermophilus及Bifidobacterium infantis發酵，期望透過微生物發酵使基質產生微生物轉化作用，生成具有免疫調節活性之醣蛋白。
首先探討不同水分含量黃豆粉中微生物生長情形，確定固態發酵之可行性。之後以 NK-92 MI 細胞為免疫調節活性測試平臺，探討不同水分含量、微生物預發酵時間及乳酸菌發酵時間等因素，對於細胞存活率之影響。實驗結果發現，以 60% 水分含量，黃豆粉經酵母菌預發酵一天後，再由乳酸菌發酵一天及三天之組別，對免疫功效提升之效果最佳，且具有此效果之生物活性成分可能為蛋白質。 本實驗進一步以陰離子交換層析分離純化粗蛋白萃取液，發現經過發酵後蛋白質及醣組成有改變之情形，將區分物以 NK-92 MI 細胞測試其免疫調節活性，結果顯示，黃豆粉經酵母菌預發酵一天後，再由乳酸菌發酵一天及三天之組別，能顯著促進 NK-92 MI 細胞增生，並增加其毒殺 K-562 細胞之活性。 | zh_TW |
| dc.description.abstract | Natural killer cells play a vital role in immune system, which is the first line defence against viral infection and in preventing the development of cancers. To modulate immune function by diets has been a topic of interest to researchers in recent years. Glycoproteins isolated from natural sources have been proven to be an immunomodulatory potentiators. Soybean contains high quality protein, moreover, it has been reported that lactic acid bacteria fermented soy-based products possesses immunomodulatory activity. However, the soy-based products fermented by lactic acid bacteria were conducted majorly by submerged fermentation.
In this study, we fermented soymeal by using two stage solid state fermentation. The soymeal inouculated Lactobacillus acidophilus, Streptococcus thermophilus and Bifidobacterium infantis after Saccharomyces cerevisiae pre-fermentation. We expected that the fermentation process could produce immunomodulatory glycoprotein due to bio-conversion of microorganism. The effect of soymeal with various moisture contents on microorganism growth was examined to evaluate feasibility of solid-state fermentation. In addition, the effect of various moisture contents, Saccharomyces cereviseae pre-fermentation time and lactic acid bacteria fermentation time on NK-92 MI cell viability were also be investigated. It was found that soymeal fermented by lactic acid bacteria for one and three days after yeast pre-fermentation for one day could promote NK-92 MI proliferation and cytotoxic activity of NK-92 MI cells against K562 cell. Furthermore, the bioactive compound is likely to be protein. The crude protein extract was fracrionated by anion-exchange chromatography. The elution profile of fermented sample was different from raw material. It was found that the fraction 4 of soymeal fermented by lactic acid bacteria for one and three days after yeast pre-fermentation for one day could promote NK-92 MI cells proliferation, and enhanced the cytotoxic activity of NK-92 MI cells against K562 cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T05:11:22Z (GMT). No. of bitstreams: 1 ntu-103-R01641007-1.pdf: 1190696 bytes, checksum: 088cdcbe8604c570e0e734e7eafa25d4 (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要 ii Abstract iii 第一章、 文獻回顧 1 第一節、 免疫系統 (Immune system) 1 一、 先天性免疫反應 (Innate or non-specific immunity) 2 (一) 物理性屏障 (Physical or structural hindrance) 2 (二) 生理性屏障 (Physiologic barriers) 2 (三) 噬菌性屏障 (Endocytic and phagocytic barriers) 2 (四) 自然殺手細胞 (Natural killer cells, NK cells) 2 二、 後天性免疫反應 (Aquired or specific immunity) 2 (一) T淋巴細胞 (T lymphocytes) 3 (二) B 淋巴細胞 (B lymphocytes) 3 (三) 細胞免疫反應 (Cellular immune response) 3 (四) 體液免疫反應 (Humoral immune response) 4 第二節、 自然殺手細胞在免疫系統扮演的角色 5 一、 自然殺手細胞的功能 6 (一) 異常細胞的消滅 6 (二) 調節造血細胞 6 (三) 骨髓移植時的抗白血病效應 6 (四) 過敏及自體免疫疾病 7 二、 自然殺手細胞的活化 7 (一) 活化及抑制型受體 7 (二) 抗體倚賴型細胞毒殺作用 (Antibody-dependent cellular cytotoxicity, ADCC) 9 (三) 細胞激素的分泌 9 三、 NK-92 MI 細胞株特性 9 第三節、 飲食對於免疫系統之影響 10 第四節、 醣蛋白 11 一、 醣蛋白簡介 11 (一) 醣蛋白的結構 11 (二) 醣蛋白的功能 11 二、 醣蛋白的免疫生物活性 12 第五節、 黃豆 13 一、 黃豆的營養成分 13 (一) 蛋白質 13 (二) 脂質 13 (三) 碳水化合物 14 (四) 皂素 (Saponins) 14 (五) 異黃酮 (Isoflavone) 14 (六) 微量元素 14 二、 黃豆發酵產品 15 第六節、 固態發酵 16 一、 固態發酵之特性 16 二、 水分含量之影響 18 第七節、 發酵菌株之挑選 19 一、 乳酸菌 19 二、 酵母菌 19 三、 共培養發酵 20 第二章、 實驗目的與設計 21 第一節、 實驗目的 21 第二節、 實驗設計 22 一、 實驗流程 23 二、 實驗架構 24 (一) 第一部分：兩階段式固態發酵可行性之評估 24 (二) 第二部分：樣品免疫調節活性之評估 25 (三) 第三部分：樣品組成變化與免疫調節活性相關性之比對 26 第三章、 實驗材料與方法 27 第一節、 兩階段式固態發酵可行性之評估 27 一、 實驗材料 27 (一) 發酵基質 27 (二) 發酵菌株 27 (三) 實驗藥品 27 二、 器材與儀器 29 三、 實驗方法 29 (一) 不同粉水比黃豆粉之配製 29 (二) 菌種之活化與保存 30 (三) 發酵樣品之製備 30 (四) 菌體生長情形之測定 31 (五) 發酵終止 31 (六) 分析方法 32 第二節、 樣品免疫調節活性之評估 34 一、 實驗材料 34 (一) 實驗細胞株 34 (二) 實驗藥品 34 二、 器材與儀器 35 三、 實驗方法 35 (一) 樣品脫脂 35 (二) 樣品萃取 35 (三) 蛋白質粗萃液之製備 36 (四) 細胞培養 36 (五) 細胞存活率 (CCK-8 assay) 37 (六) NK-92 MI毒殺 K-562 細胞實驗 38 (七) 統計分析 39 第三節、 樣品組成變化與免疫調節活性相關性之比對 40 一、 實驗藥品 40 二、 器材與儀器 40 三、 實驗方法 41 (一) 樣品製備 41 (二) 活性醣蛋白的分離純化 41 第四章、 結果與討論 43 第一節、 兩階段式固態發酵可行性之評估 43 一、 發酵條件之確立 43 (一) 不同粉水比之基質水分含量及水活性 43 (二) S.cerevisiae 於不同水含量基質下之生長情形 44 (三) 乳酸菌於酵母菌預發酵黃豆粉中之生長情形 47 (四) 發酵過程中總醣及還原糖變化情形 51 (五) 發酵過程中蛋白質及胜肽變化情形 54 第二節、 樣品免疫調節活性之評估 57 一、 評估樣品粗萃液之免疫調節活性 57 二、 評估發酵樣品蛋白質粗萃液之免疫調節活性 63 第三節、 樣品組成變化與免疫調節活性相關性之比對 69 一、 以離子交換層析法區分黃豆粗醣蛋白 69 二、 評估醣蛋白質區分物之免疫調節活性 75 第五章、 結論 79 參考文獻 80 附錄 88 | |
| dc.language.iso | zh-TW | |
| dc.title | 利用兩階段式固態發酵豆粉開發具有免疫調節活性之產品 | zh_TW |
| dc.title | Development of Fermented Soybean Meals for Modulability of Immune Response by Using Two Stage Solid State Fermentation | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 潘敏雄(Min-Hsiung Pan),潘子明(Tz-Ming Pan),柯逢年(Peng-Nisn Ke) | |
| dc.subject.keyword | 醣蛋白,免疫調節,兩階段式發酵, | zh_TW |
| dc.subject.keyword | Glycoprotein,immunomodulatory,two-stage fermentation, | en |
| dc.relation.page | 89 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-08-19 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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