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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉嚞睿(Je-Ruei Liu) | |
| dc.contributor.author | Shu-Jie Li | en |
| dc.contributor.author | 李淑婕 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:35:26Z | - |
| dc.date.available | 2023-09-01 | |
| dc.date.copyright | 2022-01-03 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-09-07 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82080 | - |
| dc.description.abstract | "近年來，由於科學證據不斷地表明益生菌對人類健康有益，有關益生菌的保健食品已經在商業市場上占據一席之地。益生菌具有免疫調節、抗發炎、抗氧化等功效，且益生菌易被攝取和其經濟價值高。本實驗針對土壤中篩選出的Bacillus sp. AC，進行基本分子與生化特性分析。經鑑定試驗結果顯示，AC屬於Bacillus velezensis。本實驗並探討AC及其産物之抗氧化活性，一方面從清除1,1-diphenyl-2-picrylhydrazyl（DPPH）自由基能力測定和清除氫氧自由基能力測定；另一方面，以人類胚肺成纖維細胞MRC-5建立早期衰老細胞模式，以過氧化氫（H2O2）誘導MRC-5使細胞衰老，並以流式細胞儀分析細胞老化週期，以及將芽孢桿菌及其產物以預處理方式在過氧化氫誘導細胞老化前給予保護作用。結果顯示，Bacillus velezensis AC其清除DPPH自由基能力和清除氫氧自由基能力在特定濃度下有顯著提升。在過氧化氫誘導氧化損傷之前，MRC-5細胞用AC或其細胞成分進行預處理。對MRC-5細胞氧化損傷主要指標為超氧化物歧化酶（superoxide dismutase, SOD）、穀胱甘肽過氧化物酶（glutathione peroxidase, GPx）等抗氧化酶的表達進行分析，結果表明其胞內液（intracellular fraction, ICF）具有較高的抗氧化酶活性。在這項研究中，我們證明 B. velezensis AC 的細胞胞內液具有顯著的自由基清除能力和抗氧化活性。通過細胞模型實驗，我們也證明 B. velezensis AC 可能是益生菌食品補充劑的候選者。綜上所述， B. velezensis AC 具有清除自由基的分子，且富含抗氧化活性，期望未來在動物實驗上能有好的表現並發展爲健康保健食品。" | zh_TW |
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| dc.description.tableofcontents | 致謝 i 中文摘要 ii Abstract iii 目錄 iv 第一章、文獻回顧 1 第一節、老化 1 一、 老化之定義 1 二、 老化學說 1 三、 氧化壓力與氧化應激 3 四、 細胞凋亡 4 五、 細胞週期 4 第二節、益生菌 9 一、 益生菌定義及鑑定 9 二、 芽孢桿菌作為益生菌之特性 10 三、 芽孢桿菌種類 12 四、 芽孢桿菌的抗氧化能力 16 第二章、材料與方法 20 第一節、 實驗架構 20 第二節、 實驗藥品與儀器 21 一、 試藥 21 二、 實驗儀器與器材 22 三、 實驗菌株 23 四、 細胞株 23 五、 培養液 24 第三節、芽胞桿菌之基本生理生化特性 25 一、 菌種活化與保存 25 二、 基本特性分析 25 三、 菌株碳源利用能力 25 四、 16S rRNA和gyrB基因定序 26 五、 親緣關係樹的建立 26 六、 生長曲線 27 七、 抗病原菌能力 27 八、 耐膽鹽試驗 28 九、 耐酸試驗 28 十、 抗生素敏感性試驗 28 十一、 芽孢桿菌之胞內萃取物與細胞壁樣品製備 29 十二、 腸道貼附 30 十三、 內毒素測定 31 第四節、Bacillus spp.的胞內液和細胞壁之抗氧化力測定 31 一、 清除DPPH自由基能力測定 31 二、 清除氫氧自由基能力測定 32 第五節、Bacillus spp.及其產物之抗氧化細胞試驗 33 一、 細胞培養 33 二、 過氧化氫誘導細胞早期衰老之測定結果 34 三、 細胞預處理以抗細胞早期衰老 35 四、 SOD活性分析 35 五、 GPx活性分析 36 六、 細胞週期檢測 37 七、 老化相關基因mRNA表現量分析 38 八、 統計分析 39 第三章、結果 46 第一節、Bacillus velezensis AC之基本生理生化特性 46 一、 菌株菌落型態觀察與革蘭氏染色 46 二、 菌株碳源利用性 46 三、 菌株分子鑑定 47 第二節、Bacillus velezensis AC 作為益生菌之潛力 48 一、 病原菌抑制能力評估 48 二、 酸耐受性試驗 48 三、 膽鹽耐受性試驗 49 四、 抗生素敏感性試驗 49 五、 腸道貼附能力評估 50 六、 内毒素测定 50 第三節、Bacillus spp.的胞內液和細胞壁之抗氧化力測定 65 一、 Bacillus spp.之胞內液和細胞壁之清除DPPH自由基能力測定結果 65 二、 Bacillus spp之胞內液和細胞壁之清除氫氧自由基能力測定結果 65 第四節、Bacillus spp. 及其産物之抗氧化及抗老化機制探討 66 一、 過氧化氫誘導細胞早期衰老之測定結果 66 二、 SOD活性分析 66 三、 GPx活性分析 67 四、 細胞週期檢測 67 五、 老化相關基因mRNA表現量分析 68 第四章 討論 86 第一節、菌種鑑定結果 86 第二節、Bacillus velezensis AC 作為益生菌之潛力 87 一、 病原菌抑制能力評估 87 二、 酸耐受性試驗 88 三、 膽鹽耐受性試驗 89 四、 抗生素敏感性試驗 90 五、 腸道貼附能力評估 91 六、 內毒素基因評估 91 第三節、Bacillus spp. 及其産物之抗氧化及抗老化機制探討 92 一、 Bacillus spp.的胞內液和細胞壁抗氧化能力測定結果 92 二、 過氧化氫誘導細胞早期衰老之測定結果 95 三、 SOD活性分析 95 四、 GPx活性分析 95 五、 細胞週期檢測 96 六、 老化相關基因mRNA表現量分析 96 第五章 結論 98 第六章 參考資料 99 | |
| dc.language.iso | zh-TW | |
| dc.subject | 抗氧化 | zh_TW |
| dc.subject | 芽孢桿菌 | zh_TW |
| dc.subject | 氧化應激 | zh_TW |
| dc.subject | 抗老化 | zh_TW |
| dc.subject | oxidative stress. | en |
| dc.subject | anti-senescence | en |
| dc.subject | antioxidant | en |
| dc.subject | Bacillus velezensis | en |
| dc.title | 貝萊斯芽孢桿菌AC菌株的鑑定及抗氧化能力分析 | zh_TW |
| dc.title | Identification of a Bacillus sp. AC strain and analysis of its antioxidant ability | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張慧雯(Hsin-Tsai Liu),謝建元(Chih-Yang Tseng),鄭永祥,劉啟德 | |
| dc.subject.keyword | 芽孢桿菌,抗氧化,抗老化,氧化應激, | zh_TW |
| dc.subject.keyword | Bacillus velezensis,oxidative stress.,antioxidant,anti-senescence, | en |
| dc.relation.page | 108 | |
| dc.identifier.doi | 10.6342/NTU202102980 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-09-08 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物科技研究所 | zh_TW |
| dc.date.embargo-lift | 2023-09-01 | - |
| 顯示於系所單位： | 生物科技研究所 | |
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