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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 徐濟泰(Jih-Tay Hsu) | |
dc.contributor.author | I-Hung Chen | en |
dc.contributor.author | 陳宜鴻 | zh_TW |
dc.date.accessioned | 2021-06-12T17:59:37Z | - |
dc.date.available | 2010-02-01 | |
dc.date.copyright | 2008-02-01 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-01-28 | |
dc.identifier.citation | 參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27268 | - |
dc.description.abstract | 本試驗針對R. albus 7所分泌細菌素之半純化條件、特性與未來用於商業飼糧之可能性進行探討。R. albus 7細菌使用不含瘤胃液之化學培養基進行培養,參考文獻添3μM 3-phenylpropanoic acid與0.2% Tween 80當為刺激R. albus 7生長之刺激原,在連續培養下,從12至32小時的培養時間每6小時收取R. albus 7細菌素一次,尋找最適生產時間點,透過快速蛋白質分析儀(FPLC)以ion-exchange進行R. albus 7細菌素的半純化。特性分析則使用細菌素對Enterobacter aerogenes (BCRC 10370)、Salmonella enterica (BCRC 10747)、Pseudomonas aeruginosa (BCRC 10944)、Bacillus subtilis (BCRC 10029)及Staphylococcus aureus (BCRC 10451)等五種指標菌之抑菌效果。細菌素分子量的分析使用10% 原態電泳膠片(native-PAGE)抑菌活性測定方法估計,最小抑菌濃度則使用微劑量液態稀釋測定法(microtitre broth dilution method)進行測定,以飼料工業打粒常用溫度65℃,進行細菌素耐熱性檢驗,而在蛋白酶耐受性部分則使用胃蛋白酶(pepsin)、胰綜合酶(pancreatin)及商業蛋白酶(prokinase)等三種蛋白酶作用與比較,最終檢測使用1% bovine serum albumin為保護劑之效果。
試驗結果顯示R. albus 7能夠使用不含瘤胃液之培養基生產細菌素,且以培養時間24小時的細菌素抑菌總活性最佳,R. albus 7之未純化細菌素在初始試驗部分,即能展現最低64.5 % 最高80.5 % 的抑菌效果,後經5 kDa濃縮膜濃縮100倍,並使用FPLC配合DEAE管柱進行層析分離,在食鹽(NaCl)濃度0.23 M下,可獲得回收率達23 % 之半純化細菌素。使用原態膠體電泳進行活性分析結果顯示,R. albus 7細菌素應為33 kDa左右的蛋白質。經耐受性試驗測試,R. albus 7細菌素不能耐受pepsin、pancreatin、protease的水解作用或65℃之加熱處理,且進行家禽體外模擬消化試驗也顯示細菌素抑菌活力明顯下降74.7%以上(P < 0.05)。但當添加1% BSA為保護劑的情況下,則可減少抑菌活力下降比率達13%(P < 0.05)。在最小抑菌濃度測試中,R. albus 7細菌素若需達成50%抑菌濃度則至少需0.75 mg/mL,90%抑菌濃度則最少為6.5 mg/mL。培養基中添加刺激原3-phenylpropanoic acid(PPA)與Tween 80,可以使R. albus 7生長提早2小時達到高峰,同時使細菌素提早2小時產出,兩者共同添加更可以提早4小時產出,產量更可達到未添加時的2.5倍,其中又以有Tween 80添加的兩種處理均能獲得較好的抑菌效果(p < 0.05)。 綜合上述,R. albus 7細菌素具備對抗常見病原菌的能力,在考量其特性後,可添加保護劑以增其耐受力,應能擴展更廣泛的應用範圍,使R. albus 7細菌素成為一具有替代抗生素潛力之產品。 | zh_TW |
dc.description.abstract | Our research was set up to find more detailed characteristics of the bacteriocin from R. albus 7 and its potential as alternative of antibiotics. We also tried to establish reliable production and semi-purification procedures. In our results, R. albus 7 could be cultured in the medium without ruminal fluid to produce bacteriocin effectively against five pathogens (Enterobacter, Bacillus, Pseudomonas, Samonella, Staphylococcus). MIC50 and MIC90 of R. albus 7 bacteriocin were as low as 0.75 mg/mL and 6.5 mg/mL, respectively. The bacteriocin is highly released after 24 hr of incubation. Inclusion of 3μM 3-phenylpropanoic acid(PPA)and 0.2 % Tween 80 in culturing medium improved 2.5 times production of bacteriocin. Native-gel electrophoresis showed that bacteriocin of R. albus 7 had a molecular weight about 33 kDa. Semi-purification procedure resulted in 23% recovery rate of bacteriocin by using the DEAE column in FPLC system. The bacteriocin of R. albus 7 could be destroyed by pepsin, protease, and pancreatin. It also could be inactivated after heating at 65℃for 1 hr. Simulated in vitro avian digestion decreased the antagonistic activity by 74.7%. But, addition 1% BSA could retain 13% more antagonistic activity.
In conclusion, the bacteriocin of R. albus 7 is a heat-labile small protein. PPA and Tween 80 are recommended to be included in its production culture medium. DEAE column was suitable for its semi-purification. The bacteriocin of R. albus 7 has the chance as an antibiotic alternative with protectant added. | en |
dc.description.provenance | Made available in DSpace on 2021-06-12T17:59:37Z (GMT). No. of bitstreams: 1 ntu-97-R94626006-1.pdf: 957270 bytes, checksum: 4771ea1a23503eecd0f4c7805c5e9967 (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 目錄 頁次
致謝 i 目錄 iii 表次 iv 圖次 vi 壹、 前言 1 貳、 中文摘要 2 參、 英文摘要 4 肆、 文獻檢討 6 伍、 材料與方法 27 陸、 結果與討論 55 柒、 結論 83 捌、 參考文獻 84 玖、 附錄 92 | |
dc.language.iso | zh-TW | |
dc.title | 瘤胃細菌Ruminococcus albus 7細菌素的半純化與特性分析 | zh_TW |
dc.title | Semi-purification and characterization of bacteriocin from Ruminococcus albus 7 | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 魏恆巍(Wei-Heng Wei),劉?睿(Jhe-Ruei Liou),王翰聰(Han-Tsung Wang) | |
dc.subject.keyword | 細菌素,瘤胃細菌,抗生素替代, | zh_TW |
dc.subject.keyword | bacteriocin,ruminal bacteria,antibiotic alternative, | en |
dc.relation.page | 94 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-01-29 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
顯示於系所單位: | 動物科學技術學系 |
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