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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 潘子明 | |
dc.contributor.author | Tzu-Hsing Lin | en |
dc.contributor.author | 林姿杏 | zh_TW |
dc.date.accessioned | 2021-06-16T03:42:57Z | - |
dc.date.available | 2018-03-13 | |
dc.date.copyright | 2015-03-13 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-02-11 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/54967 | - |
dc.description.abstract | 乳酸菌 (lactic acid bacteria, LAB) 廣泛存在自然界,泛指一群能發酵碳水化合物產生多量乳酸的細菌。乳酸菌製作的發酵食品及發酵飲品食用歷史悠久,原本是為食品保存,後來漸漸發展成地區特有的傳統發酵食品。乳酸菌的代謝產物包括有機酸 (乳酸與醋酸)、二乙醯、過氧化氫、細菌素與低分子量的化合物等,這些具有抗菌能力的代謝產物是延長發酵食品保存時間的關鍵。這些乳酸菌代謝產物中,最具抑菌活性的是由細菌產生具有抑菌或殺菌活性的胜肽或小分子化合物。在目前的研究中,學者對其抑菌能力的廣效性及作為天然食品添加物延長食品保存期限最感興趣。本研究室先前的研究結果顯示,分離的乳酸菌株 Lactobacillus paracasei subsp. paracasei NTU 101 與Lactobacillus plantarum NTU 102 具有廣效的抑菌範圍。本研究評估此二株乳酸菌上清液純化所得抗菌活性物質之抗菌範圍及活性,並以核磁共振光譜儀分析其結構。以誘導齲齒之動物模式測試抗菌物質是否具抗齲能力,結果顯示 L. paracasei subsp. paracasei NTU 101 及 L. plantarum NTU 102 上清液所含抗菌物質可以抑制大鼠口腔中變形鏈球菌菌數增長並降低齲齒指數,與未塗抹抗菌物質的組別有顯著差異 (p < 0.05),顯示其良好抗齲能力,可做為開發口腔保健產品之開發素材。為提升其應用的可能性,另以反應曲面法探討最適替代培養基,以豆漿與糖蜜為替代培養基可大幅降低生產成本,如要應用於大規模商業生產,仍須提升產量以降低耗能與生產成本。 | zh_TW |
dc.description.abstract | Lactic acid bacteria (LAB) are widely distributed in nature such as in dairy, fish as well as vegetables and grains. LAB fermented foods are less susceptible to microbial spoilage and prolong shelf life in food preservation. Previous studies showed that metabolites produced from LAB including organic acids (lactic acid, acetic acid), diacetyl, hydrogen peroxide, bacteriocins and some low molecular weight compounds have antimicrobial abilities. Among these antimicrobial metabolites, peptides or the low molecular weight compounds with bacteriostatic and bactericidal action, attracted interest of researchers and food producers for the potential to be biopreservative additives. In this study, we investigated the antimicrobial activity of supernatant of Lactobacillus paracasei subsp. paracasei NTU 101 and Lactobacillus plantarum NTU 102 and demonstrated the broad inhibitory spectrum. The antimicrobial substances were purified, characterized and identified by nuclear magnetic resonance (NMR) techniques. In animal experiments, we assessed Streptococcus mutans counts and caries scores to evaluate the decay process during a 68-day period. These agents significantly reduced oral cariogenic bacteria and caries scores (p < 0.05). The results revealed that the bacteriological factors involved in the caries process can be suppressed. These results suggested that the antimicrobial substances produced by L. paracasei subsp. paracasei NTU 101 and L. plantarum NTU 102 have protective functions against dental caries development. The production of antimicrobial substances was optimized by the Box-Behnken experimental design. An empirical model was developed through response surface methodology to describe the relationship between variables (molasses, soy-milk, and incubation temperature). The results revealed that an alternative medium could be used for large-scale commercial production of the antimicrobial substances, with potential prospects for application. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T03:42:57Z (GMT). No. of bitstreams: 1 ntu-104-D97b47101-1.pdf: 27377042 bytes, checksum: 983b4666d2eaef7193914f714037d340 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 縮寫表 iv
摘要 x Abstract xi 第一章 研究背景 1 一、乳酸菌 (lactic acid bacteria) 1 (一) 乳酸菌之簡介 1 (二) 乳酸菌之營養機能及健康效益 1 (三) 益生菌 (probiotics) 之簡介 6 二、齲齒 (dental caries) 11 (一) 齲齒之簡介 11 (二) 齲齒的成因 15 (三) 致齲細菌 (cariogenic bacteria) 17 (四) 變形鏈球菌之毒力因子 (virulence factors) 18 (五) 變形鏈球菌致齲機制 23 三、益生菌與齲齒 27 四、研究動機與目的 29 五、研究架構 31 第二章 開發大量生產抗菌物質之替代培養基 32 一、前言 32 二、材料與方法 33 (一) 試驗菌株 33 (二) 菌種活化與保存 33 (三) 抗菌活性測試 33 (四) 選擇替代培養基組成 34 (五) 以反應曲面法探討生產抗菌物質的最適條件 35 三、結果 36 (一) 反應曲面法中心點參數選擇 36 (二) 以反應曲面法進行培養條件最適化 41 四、討論 52 第三章 乳酸菌抗菌物質對變形鏈球菌之抑制作用 53 一、前言 53 二、材料與方法 53 (一) 試驗菌株 53 (二) 製備乳酸菌上清液 54 (三) 萃取抗菌物質 54 (四) 抗菌物質對指標菌株之影響 55 (五) 變形鏈球菌對羥基磷灰石的降解能力 55 (六) 抗菌物質對致齲性的影響 56 (1) 變形鏈球菌產酸能力 56 (2) 變形鏈球菌合成多醣量 56 (3) 變形鏈球菌菌體表面的疏水性 56 (4) 生物膜 57 (5) 黏附性 57 1. 抽取總RNA 58 2. RNA 品質確認及濃度測定 58 3. 製備 cDNA 59 4. RT-PCR 分析條件 59 三、結果 61 (一) 抗菌物質對指標菌株之影響及對羥基磷灰石的降解能力 61 (二) 抗菌物質對致齲性的影響 61 (1) 產酸能力 61 (2) 抗菌物質對合成多醣量之影響 64 (3) 抗菌物質對菌體表面疏水性之影響 64 (4) 抗菌物質對變形鏈球菌生物膜生合成相關基因表現量之影響 67 四、討論 72 第四章 乳酸菌發酵產物對變形鏈球菌之抗齲作用 75 一、前言 75 二、材料與方法 76 (一) 實驗動物飼養與照料 76 (二) 樣品製備 77 (三) 誘導齲齒 77 (四) 動物犧牲及切斷顎骨 (mandible) 78 (1) 動物犧牲 (sacrifice) 78 (2) 齲齒指數 78 三、結果 82 (一) 變形鏈球菌菌數 82 (二) 齲齒指數 90 四、討論 93 第五章 乳酸菌所生產抗菌物質之純化 94 一、前言 94 二、材料與方法 94 (一) 試驗菌株 94 (二) 菌種活化與保存 96 (三) 抗菌活性測試 96 (1) 有機酸排除試驗 96 (2) 蛋白酶敏感性測試 96 (四) pH 值及溫度敏感性測試 97 (五) 抗菌物質萃取 97 (1) 有機溶劑粗萃取 97 (2) 管柱層析 (column chromatography) 97 (3) 高效液相層析 (high performance liquid chromatography, HPLC) 98 (六) Nuclear Magnetic Resonance Spectroscopy (NMR) 分析鑑定結構式 98 (七) 最低抑菌濃度 (minimum inhibitory concentration, MIC) 測定 99 三、結果 100 (一) 抗菌活性測試 100 (二) 抗菌物質的萃取與分離 105 (三) 最低抑菌濃度 (minimum inhibitory concentration, MIC) 116 四、討論 118 結論 120 參考文獻 121 附錄 137 | |
dc.language.iso | zh-TW | |
dc.title | 本土乳酸菌 Lactobacillus paracasei subsp. paracasei NTU 101 及 Lactobacillus plantarum NTU 102 所生產抗菌物質之純化與抗齲應用 | zh_TW |
dc.title | Purification and application of anticarious substances produced by Lactobacillus paracasei subsp. paracasei NTU 101 and Lactobacillus plantarum NTU 102 | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-1 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 周正俊,廖啟成,陳明汝,邱秋霞,蔡宗佑 | |
dc.subject.keyword | 乳酸菌,抗菌活性,齲齒,抗齲能力,反應曲面法,最適化, | zh_TW |
dc.subject.keyword | lactic acid bacteria,antimicrobial activity,dental caries,anticaries,response surface methodology,optimization, | en |
dc.relation.page | 153 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-02-11 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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