菌株TKU037係篩選自淡水土壤，液態發酵NB (Nutrient Broth) 能生產α-葡萄糖苷酶抑制劑和抗菌物質，經鑑定係屬類芽胞桿菌（Paenibacillus sp.）之一株細菌。本研究探討Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之較適條件，以及所生產α-葡萄糖苷酶抑制劑之pH值與熱安定性，並且分離出具α-葡萄糖苷酶抑制活性之一化合物1-methylhydantoin。

Bacterial strain TKU037 produced α-glucosidase inhibitors and antibacterial materials when cultured in nutrient broth (NB). Strain TKU037 was isolated from Taiwanese soils and identified as Paenibacillus species. Optimum culture condition for the production of α-glucosidase inhibitors and the pH and thermal stability of the produced inhibitors have been studied. A compound (1-methylhydantoin) with α-glucosidase inhibitor activity has been isolated from the culture supernatant.

中文摘要	I
英文摘要	II
目錄	III
圖目錄	VII
表目錄	IX

第一章 緒論	1
第二章 文獻回顧	3
2.1 實驗菌種Paenibacillus sp.	3
2.2 α-葡萄糖苷酶 (α-glucosidase)	3
2.3 α-葡萄糖苷酶抑制劑 (α-glucosidase inhibitor)	4
第三章 材料與方法	7
3.1 菌株	7
3.2 化學材料	7
3.3 實驗儀器	8
3.4 生產菌株之篩選與分離	9
3.5 α-葡萄糖苷酶抑制活性之測定	9
3.6 計算公式算法	10
3.6.1 抑制率	10
3.6.2 抑制劑濃度U(unit)/mL	10
3.7 α-葡萄糖苷酶抑制劑較適生長條件探討	10
3.7.1 碳/氮源的影響	10
3.7.2 培養曝氣量對α-葡萄糖苷酶酶抑制率之影響	11
3.7.3 培養基質濃度對α-葡萄糖苷酶酶抑制率之影響	11
3.7.4 培養溫度對α-葡萄糖苷酶酶抑制率之影響	11
3.8 α-葡萄糖苷酶抑制劑之分離與純化	12
3.8.1 粗抑制劑之製備	12
3.8.2 離子交換樹酯層析	12
3.8.3 十八烷基矽烷(ODS;Octadecylsilane)膠體管柱層析	12
3.8.4 矽膠(Silica gel) 管柱層析	13
3.9 α-葡萄糖苷酶抑制劑之安定性	13
3.9.1 抑制劑熱安定性	13
3.9.2 抑制劑pH值安定性	13
3.9.3抑制劑有機溶液安定性	14
3.10 α-葡萄糖苷酶抑制劑特性之分析	14
3.10.1 TLC (Thin Layer Chromatography)	14
3.10.2 HPLC (High Performance Liquid Chromatography)	14
3.11 Paenibacillus sp. TKU037之抗菌特性	15
第四章 結果與討論	16
4.1 α-葡萄糖苷酶抑制劑生產菌株之篩選鑑定	16
4.2.1 較適碳/氮源之種類	16
4.2.2 較適培養曝氣量	18
4.2.3 較適培養基質濃度	19
4.2.4 較適培養溫度	20
4.3 α-葡萄糖苷酶抑制之分離純化	21
4.3.1 離子交換樹酯層析	22
4.3.2 十八烷基矽烷(ODS;Octadecylsilane)膠體管柱層析	22
4.3.3 矽膠(Silica gel) 管柱層析	23
4.4 α-葡萄糖苷酶抑制之定性測試	25
4.4.1 抑制劑熱安定性	25
4.4.2 抑制劑pH值安定性	27
4.4.3抑制劑有機溶液安定性	29
4.5 α-葡萄糖苷酶抑制劑特性之分析	31
4.5.1 TLC(thin layer chromatography)	31
4.5.2 HPLC (high performance liquid chromatography)	33
4.6 Paenibacillus sp. TKU037之抗菌特性	34
第五章 結論	35
第六章 參考文獻	37


 

圖目錄
圖4.1 不同碳/氮源對Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之影響	17
圖4.2 不同培養曝氣量對Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之影響	18
圖4.3 不同培養基質濃度對Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之影響	19
圖4.4 不同培養溫度對Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之影響	20
圖4.5 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之純化流程圖	21
圖4.6 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之熱安定性	26
圖4.7 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之pH值安定性	28
圖4.8 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑之有機溶劑安定性	30
圖4.9 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑純化之TLC圖	32
圖4.10 Paenibacillus sp. TKU037生產α-葡萄糖苷酶抑制劑純化之HPLC圖	33

表目錄
表2.1 糖尿病藥物成分比較	5
表2.2 α-葡萄糖苷酶抑制劑成分比較	6
表3.1 化學材料縮寫表	8
表4.1 Paenibacillus sp. TKU037α-葡萄糖苷酶抑制劑純化表	24
表4.2 Paenibacillus sp. TKU037之抗菌表	34

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