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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25522完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃健雄 | |
| dc.contributor.author | Wei-Ying Tseng | en |
| dc.contributor.author | 曾瑋盈 | zh_TW |
| dc.date.accessioned | 2021-06-08T06:17:05Z | - |
| dc.date.copyright | 2007-02-01 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-01-25 | |
| dc.identifier.citation | 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25522 | - |
| dc.description.abstract | 枯草桿菌 Bacillus subtilis 廣泛存在於自然環境中，被認定是 Generally Recognized As Safe (GRAS) 菌種，在細菌素相關研究中視為具有相當發展潛力。本研究室自淡水金沙灣沙灘篩選出菌株 Bacillus subtilis strain 6-1-1，為一具有抗菌活性之細菌菌株。B. subtilis strain 6-1-1 在平板培養基上能抑制格蘭氏陽性菌：Bacillus subtilis ATCC 6633、Micrococcus luteus BCRC 10449與Staphylococcus aureus BCRC 10541；以及格蘭氏陰性菌：Aeromonas hydrophila CKH-29 與 Escherichia coli BCRC 10450。在本研究中發現，B. subtilis strain 6-1-1 抗菌物質存在於培養離心上清液中，加入蛋白酶 trypsin、pepsin 與 proteinase K 能使其抗菌活性喪失，證明該抗菌物質應為蛋白質。含抗菌物質之培養上清液經測試對於溫度與 pH 值有很好的穩定性，80oC 下加熱處理 3 小時、pH 2 到 10 儲放四天都不致使抗菌活性消失，低溫 (4oC 或 -30oC) 下長時間儲存也能維持其抗菌活性。B. subtilis strain 6-1-1 培養液經離心、硫酸銨沉澱，與膠體過濾法等純化步驟後，可得到具抗菌活性之回收純化樣品。小分子電泳 tricine SDS-PAGE 分析純化樣品，可知 strain 6-1-1 產生之抗菌物質分子量小於 3.5 kDa，純化後純度約為 97%。將純化後抗菌物質與指標菌株共培養後，會造成指標菌種細胞死亡，推測 strain 6-1-1 所產生之抗菌物質進行殺菌作用。 | zh_TW |
| dc.description.abstract | Bacillus subtilis strain 6-1-1 was isolated from the sandy beach in Jinshawan Beach Park in Taipei County in Taiwan and was screened for its antimicrobial activity. B. subtilis strain 6-1-1, which had the broader spectrum of antibacterial activity, was active against the Gram-positive bacteria Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus; and against the Gram-negative bacteria Aeromonas hydrophila and Escherichia coli. The antimicrobial compound produced by strain 6-1-1 existed in the cell-free culture supernatant, and was determined as a proteinaceous substance, as it was sensitive to protease. It remained its antimicrobial activity under heat treatment and long-term storage at low temperature. The antimicrobial activity was stable between pH 2 and 10. The bacteriocin-like substance produced by strain 6-1-1 was purified sequentially by centrifugation, ammonium sulfate precipitation, and gel filtration. The molecular weight was determined by tricine SDS-PAGE to be about below 3.5 kDa. And the bacteriocin-like substance produced by strain 6-1-1 acts as a bacteriocidal action. B. subtilis strain 6-1-1 produces a low molecular mass bacteriocin-like compound, with a wide spectrum of activity, and potential technological properties in thermostability and pH stability for use as a bio-preservative. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T06:17:05Z (GMT). No. of bitstreams: 1 ntu-96-R92b47106-1.pdf: 1013938 bytes, checksum: 1085e1aaf599293cde7336e71508c90f (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 第一章 前言 …………………………………………………………………………1
第二章 文獻回顧 ……………………………………………………………………2 第一節 抗微生物藥物發展趨勢 ………………………………………………2 第二節 抗微生物胜肽 …………………………………………………………3 第三節 細菌素的發現與定義 …………………………………………………5 第四節 細菌素與抗生素 ………………………………………………………5 第五節 細菌素的分類 …………………………………………………………7 第六節 細菌素的作用機制 ……………………………………………………7 第七節 細菌素的利用 ………………………………………………………12 7.1 腸道菌 ……………………………………………………………………12 7.2 乳酸菌 ……………………………………………………………………13 7.3 芽胞桿菌屬 ………………………………………………………………15 第八節 細菌素的純化與鑑定 ………………………………………………17 第三章 研究動機、目的與實驗架構 ……………………………………………20 第一節 研究動機與目的 ……………………………………………………20 第二節 實驗架構 ……………………………………………………………21 第四章 材料與方法 ………………………………………………………………22 第一節 菌種與培養條件 ……………………………………………………22 1.1 指標菌種 …………………………………………………………………22 1.2 使用菌種 …………………………………………………………………22 第二節 藥品與試劑 ………………………………………………………… 23 第三節 儀器與設備 ………………………………………………………… 23 第四節 一般分析方法 ……………………………………………………… 24 4.1 菌體濃度測定 ………………………………………………………24 4.2 蛋白質濃度測定 ……………………………………………………24 第五節 抗菌活性分析方法 …………………………………………………24 5.1 塗佈法 ………………………………………………………………25 5.2 點菌法 ………………………………………………………………25 5.3 擴散法 ………………………………………………………………26 5.4 96孔盤分析方法 ……………………………………………………26 第六節 含抗菌物質培養上清液分析 ………………………………………27 6.1 含抗菌物質培養上清液製備 ………………………………………27 6.2 溫度穩定性 …………………………………………………………27 6.3 pH 值穩定性 ………………………………………………………27 6.4 儲存穩定性 …………………………………………………………27 6.5 蛋白酶敏感性 ………………………………………………………28 第七節 抗菌物質分離 ………………………………………………………28 7.1 含抗菌物質培養上清液製備 ………………………………………28 7.2 硫酸銨沉澱分劃法 …………………………………………………28 7.3 膠體過濾法 …………………………………………………………29 第八節 抗菌物質分子量決定 ………………………………………………30 8.1 電泳檢定方法 ………………………………………………………30 8.2 電泳膠片之抗菌活性分析 …………………………………………31 第九節 抗菌模式 ……………………………………………………………31 第十節 抗菌範圍 ……………………………………………………………32 第五章 結果與討論 ………………………………………………………………33 第一節 抗菌範圍、抗菌活性分析方法與指標菌種的選用 …………………33 1.1 菌株 6-1-1 之抗菌範圍 ……………………………………………33 1.2 抗菌活性分析方法與指標菌種 ……………………………………33 第二節 培養上清液抗菌活性之性質分析 …………………………………36 2.1 抗菌物質存在位置 …………………………………………………36 2.2 溫度穩定性 …………………………………………………………36 2.3 pH 值穩定性 ……………………………………………………… 36 2.4 儲存穩定性 ………………………………………………………… 40 2.5 蛋白酶敏感性 ……………………………………………………… 40 2.6 討論 …………………………………………………………………40 第三節 抗菌物質之分離純化 ……………………………………………… 45 3.1 抗菌物質粗抽取與硫酸銨分劃 …………………………………… 45 3.2 膠體過濾層析法 …………………………………………………… 48 3.3 討論 ………………………………………………………………… 48 第四節 抗菌物質分子量決定 …………………………………………… 52 第五節 抗菌物質之可能作用機制 ………………………………………… 56 第六節 純化後樣品之抗菌範圍 …………………………………………… 60 第六章 結論 ……………………………………………………………………… 62 第七章 參考文獻 ………………………………………………………………… 64 | |
| dc.language.iso | zh-TW | |
| dc.subject | Bacillus subtilis | en |
| dc.subject | bacteriocin | en |
| dc.subject | antimicrobial peptide | en |
| dc.title | 一新分離枯草菌所產生細菌素之部分特性鑑定 | zh_TW |
| dc.title | Partial Characterization of Bacteriocin by a Newly Isolated Bacillus subtilis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李敏雄,潘子明 | |
| dc.subject.keyword | 枯草桿菌,抗微生物胜肽,細菌素, | zh_TW |
| dc.subject.keyword | Bacillus subtilis,antimicrobial peptide,bacteriocin, | en |
| dc.relation.page | 71 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2007-01-26 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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