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系統識別號 U0002-2007201019455800
中文論文名稱 Burkholderia cepacia TKU026 以烏賊軟骨粉為唯一碳/氮源發酵生產酪胺酸酶抑制劑之研究
英文論文名稱 The study on the tyrosinase inhibitor produced by Burkholderia cepacia TKU026 using squid pen as thesole carbon/nitrogen sources
校院名稱 淡江大學
系所名稱(中) 生命科學研究所碩士班
系所名稱(英) Graduate Institute of Life Sciences
學年度 98
學期 2
出版年 99
研究生中文姓名 許家豪
研究生英文姓名 Chia-Hao Hsu
學號 697180353
學位類別 碩士
語文別 中文
口試日期 2010-07-08
論文頁數 53頁
口試委員 指導教授-王三郎
委員-王全祿
委員-郭曜豪
中文關鍵字 酪胺酸酶抑制劑 
英文關鍵字 Burkholderia cepacia  tyrosinase inhibitor 
學科別分類 學科別醫學與生命科學生物學
中文摘要 本研究係以水產廢棄物為主要碳/氮源,經微生物發酵後其可生產酪胺酸酶抑制劑,並探討其較適培養條件為研究目的。使用菌株為Burkholderia cepacia TKU026,源自台灣北部土壤。
以50mL液態培養基含1% 烏賊軟骨粉(squid pen powder;SPP)、0.1% K2HPO4及0.05% MgSO4.7H2O於250mL三角錐瓶,置放37℃、150rpm震盪培養三天可得較高酪胺酸酶抑制劑活性(5000U/mL)。
在抗菌實驗方面發現其發酵上清液在濃縮10倍及8倍時可有效抑制Aspergillus fumigatus及Fusarium oxysporum的生長。
英文摘要 The purpose of this study is utilized of marine wastes by microbes to produce tyrosinase inhibitor. To found the optimal conditions for production of tyrosinase inhibitor were investigated. Burkholderia cepacia TKU026 was isolated from the soil in northern of Taiwan.
The optimum culture condition for tyrosinase inhibitory production were found when the culture was shaken at 37℃ for 3 days in 50 mL and 100 mL medium containing 0.1% K2HPO4, 0.05% MgSO4.7H2O and 1% squid pen powder.
Both the culture supernatant concentrated 10 and 8 times showed inhibitory activities on Aspergillus fumigates and Fusarium oxysporum.
論文目次 封面內頁
授權書
簽名頁
中文摘要 ……………………………………………………I
英文摘要………………………………………………………II
誌謝…………………………………………………………III
目錄 …………………………………………………………V
圖目錄………………………………………………………VIII
表目錄………………………………………………………IX

第一章 緒論………………………………………………1
第二章 文獻回顧…………………………………………2
2.1菌種介紹………………………………………2
2.1.1 Burkholderia cepacia TKU026 之簡介...2
2.2 酪胺酸酶(Tyrosinase) ………………………4
2.2.1酪胺酸脢之簡介………………………4
2.2.2 酪胺酸酶(Tyrosinase)的轉移途徑……5
2.3 黑色素的形成…………………………………9
2.3.1 黑色素細胞……………………………9
2.3.2 黑色素生成途徑………………………9
2.4影響黑色素生成之因素………………………9
2.5 黑色素之類型………………………………13
2.6 酪胺酸酶抑制劑(Tyrosinase inhibitor) ……13第三章 材料與方法…………………………………………17
3.1 菌株…………………………………………17
3.2 化學材料……………………………………17
3.3實驗儀器………………………………………18
3.4 生產菌株之篩選與分離……………………18
3.5 酪胺酸酶抑制率之測定……………………19
3.6 抑制率算法…………………………………19
3.7 酪胺酸酶抑制劑較適生長條件探討………19
3.7.1 碳/氮源的影響………………………19
3.7.2 烏賊軟骨粉與蝦頭粉濃度對酪胺酸酶抑制率之影響………………………20
3.7.3以其他水產廢棄物為培養基對酪胺酸酶抑制率之影響…………………………20
3.7.4 培養溫度之影響……………………20
3.7.5培養天數之影響………………………20
3.7.6培養基酸鹼值之影響…………………21
3.7.7 曝氣量對酪胺酸酶抑制率之影響……21
3.8抗黴菌試驗……………………………………21
第四章 結果與討論………………………………………22
4.1 酪胺酸酶抑制劑生產菌之篩選……………22
4.2酪胺酸酶抑制劑生產菌株之鑑定……………22
4.3較適培養條件探討……………………………26
4.3.1碳/氮源之選擇…………………………26
4.3.2 碳/氮源濃度之影響……………………27
4.3.3培養溫度之影響…………………………27
4.3.4培養基酸鹼值之影響……………………27
4.3.5培養體積之影響…………………………27
4.3.6較適培養條件整理………………………28
4.4以不同水產廢棄物為碳/氮源之比較………37
4.5 B.cepacia TKU026之抗菌特性………………40
第五章 結論…………………………………………43
參考文獻…………………………………………………44

表目錄
頁次
表 2.1 酪胺酸酶之作用…………………………………………6
表2.2 常見之酪胺酸酶抑制劑…………………………………15
表2.3 各個來源之酪胺酸酶抑制劑………………………………16
表 4.1 16SrDNA 部分序列比對結果……………………………29
表 4.2 不同酪胺酸酶抑制劑生產菌所使用之培養基……………30
表 4.3 不同SPP 濃度對酪胺酸酶抑制效果之影響………………31
表 4.4 不同碳/氮源之比較………………………………………38
表 4.5 TKU026 發酵上清液濃縮不同倍率對FO 及AF 生長之抑制效
果…………………………………………………………41

圖目錄
頁次
圖2.1 酪胺酸酶於黑色素生成圖徑中之催化位置…………………7
圖2.2 黑色素的轉移圖徑…………………………………………8
圖2.3 cAMP 經不同訊號傳遞調控黑色素的生成途徑………………12
圖4.1 Burkholderia cepacia TKU026 之顯微照片…………………23
圖4.2 Burkholderia cepacia TKU026 之16SrDNA 部分序列………24
圖4.3 16SrDNA 部分鹼基序列分析及API 試驗…………………25
圖4.4 不同碳/氮源對TKU026發酵上清液對於酪胺酸酶活性抑制之影
響…………………………………………………………32
圖4.5 TKU0265 之培養溫度對酪胺酸酶抑制效果之影響…………33
圖4.6 培養基酸鹼值對酪胺酸酶抑制效果之影響…………………34
圖4.7 培養體積對酪胺酸酶抑制效果之影響……………………35
圖4.8 Burkholderia spp.TKU026 所生產酪胺酸酶抑制劑對酪胺酸酶
之抑制效果及生長曲線……………………………………36
圖4.9 TKU026 之不同碳/氮源對酪胺酸酶抑制效果及生長曲線…39
圖 4.10 濃縮不同濃度之上清液對FO及AF生長抑制效果…………42

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