Burkholderia cepacia TKU026為以烏賊軟骨為唯一碳/氮源，篩選自台北市內湖土壤之酪胺酸酶抑制劑生產菌。本研究針對此菌發酵所得上清液之果蠅致死率以及酪胺酸酶抑制劑活性測試。果蠅外殼所含幾丁質係幾丁質合成酶所生成，而文獻報導幾丁質酶抑制劑和酪胺酸酶抑制劑具類似性質，因而期待藉由酪胺酸酶抑制劑抑制果蠅外殼幾丁質之形成而達到殺蟲效果。
  結果顯示較適培養條件為，將菌株B. cepacia TKU026接種於50 mL含有1 %烏賊軟骨粉、0.1 % K2HPO4及0.05 % MgSO4．7H2O (pH 7) 之液體培養基（裝填於250 mL震盪三角瓶加上通氣栓）於37℃經搖瓶(150 rpm) 培養2天，可具有較佳殺果蠅和酪胺酸酶抑制效果，分別為81%和95%。本實驗同時探討此殺蟲劑和酪胺酸酶抑制劑之性質與分離純化，結果指出，此物質應該為小分子化合物，具有耐熱、穩定性高的特性。

Burkholderia cepacia TKU026, a tyrosinase inhibitor-producing strain, was isolated from Taipei soil using squid pen as the sole carbon/nitrogen source. B. cepacia produced higher inhibitory activity (95%) when the organism was incubated in a baffle-based flask containing 50 mL medium of 1% squid pen powder, 0.1 % K2HPO4, and 0.05 % MgSO4 ‧7H2O (pH 7) at 37℃ for two days.
Some tyrosinase inhibitors with chitinase inhibitory activities have been investigated their insecticidal activities. In the present study, the use of tyrosinase inhibitor as insecticide was also studied. Results showed that Drosophila larval motility of the tyrosinase inhibitor was 81%. Based on the results of dialysis and heat-treatment, the inhibitor was inferred to be a thermostable compound of low molecule.

中文摘要	I
英文摘要	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 Burkholderia cepacia	2
2.2 烏賊軟骨等水產廢棄物之應用	3
2.3 殺蟲劑簡介	3
2.4 酪胺酸酶	5
2.5 酪胺酸酶抑制劑	6
2.6 幾丁質	7
2.7 幾丁質合成酶	8
2.8 幾丁質合成酶抑制劑	9
第三章	材料與方法	18
3.1 菌株	18
3.2 實驗材料	18
3.3 實驗儀器	18
3.4 生產菌株之篩選與分離	19
3.5 果蠅致死率較適培養條件探討	20
3.6 破菌方法	20
3.7 果蠅殺蟲活性測試	20
3.8 果蠅培養基	21
3.9 熱安定性處理	21
3.10 HPLC分離	22
3.11 透析處理	24
3.12 硫酸銨沉澱實驗	24
3.13 萃取	25
3.14 酪胺酸酶抑制劑測試	25
3.15 酪胺酸酶抑制率算法	25
第四章	結果與討論	26
4.1 殺蟲活性和酪胺酸酶抑制劑的不同培養天數之效果	26
4.2 熱安定性測試	27
4.3 透析實驗	29
4.4 硫酸銨沉澱結果	30
4.5 分離純化	30
第五章	結論	36
附錄	37
參考文獻	52

圖目錄
圖2.1 黑色素生合成路徑圖11
圖 2.2 麴酸、對苯二酚和熊果素之化學結構式12
圖 2.3 幾丁質合成機制13
圖 2.4 多氧霉素、尼克霉素和UDP-GlcNAc結構比較圖14
圖 2.5 幾丁質和成酶的雙活化區作用機制15
圖4.1 不同部位的殺蟲劑熱安定性結果27
圖4.2 不同加熱時間的殺蟲劑熱安定性結果28
圖4.3–4.17 附錄37

表目錄
表2.1 酪胺酸酶功能16
表2.2 各種酪胺酸酶抑制劑17
表4.1 不同天數的培養結果26
表4.2 透析結果比較29
表4.3 硫酸銨沉澱結果30

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