Serratia marcescens TKU011 係一株以蝦頭殼粉末作為唯一碳/氮源
之蛋白酶及幾丁聚醣酶生產菌。本研究發現以烏賊軟骨粉末（SPP）作
為碳/氮源時具有可生產紅色色素-靈菌紅素（prodigiosin）的能力，菌株
醱酵產生之紅色色素經分離純化後，藉由1H-NMR 及ESI-MS 確認合成
之靈菌紅素屬於prodigiosin。TKU011 生產靈菌紅素之較適培養為含有
1.5%烏賊軟骨粉末、0.1% K2HPO4 及0.05% MgSO4〃7H2O 之50 mL 液
態培養基（pH 7~9）於30℃振盪培養一天後接著移到25℃繼續培養兩
天，靈菌紅素產量可達0.978 mg/mL。醱酵上清液經試驗推測亦含有脂
胜肽類生物界面活性劑沙雷濕潤素（serrawettin），表面張力值隨著靈菌
紅素的產量增加而降低。

A protease- and chitosanase-producing strain, Serratia marcescens TKU011, was cultivated by using shrimp-head powder（SHP）as the sole carbon/nitrogen source. In this study, a prodigiosin-like pigment was produced from TKU011 by using squid pen powder（SPP）as the sole carbon/nitrogen source. prodigiosin（PG）is one of prodigiosin-like pigments produced by S.marcescens TKU011 and was purified and characterized by1H-NMR and ESI-MS to determine its chemical structure and molecular weight. The optimized conditions for prodigiosin production were cells used to inoculate in 50mL of medium containing 1.5% SPP, 0.1% K2HPO4, 0.05% MgSO4．7H2O（pH7~9）shaken at 30℃ for one day and then cultivated at 25℃ for two days. The production of PG reached 0.978 mg/mL. Besides, TKU011 culture supernatant also contained the lipopeptide biosurfactant（serrawettin）, and the production of serrawettin increased with PG production.

中文摘要	I
英文摘要	II
目錄	III
圖目錄	VII
表目錄	VIII
第一章 緒論.......................................1
第二章 文獻整理...................................2
2.1 Serratia marcescens 之分布與起源..............2
2.2 Serratia marcescens 之基本生化學特性..........2
2.3 Serratia marcescens 之應用....................3
2.3.1 幾丁質酶 (chitinase) 之生產.................3
2.3.2 蛋白酶之生產	................................4
2.3.3 色素之生產..................................4
2.4 靈菌紅素之化學結構............................5
2.5 Prodigiosin 之一般特性........................7
2.6 Prodigiosin 在細胞中之位置....................7
2.7 Prodigiosin 之培養條件與生產研究..............7
2.7.1 培養條件對 prodigiosin 生成之影響...........7
2.7.2. 大量生產 prodigiosin 之研究...............10
2.8 Prodigiosin之應用............................11
2.8.1 Prodigiosin之抗癌效果......................11
2.8.2 抗生素.....................................11
2.8.3 免疫抑制劑.................................12
2.8.4 染料.......................................12
第三章 材料與方法................................13
3.1 實驗架構.....................................13
3.2 實驗菌株.....................................14
3.3 實驗材料.....................................14
3.4 實驗儀器.....................................15
3.5 碳氮源種類篩選...............................15
3.6  Prodigiosin 之定量..........................16
3.7  Prodigiosin 產量之計算方式..................16
3.8  Prodigiosin 之純化..........................17
3.9 較適培養條件探討.............................18
3.9.1 培養基酸鹼值...............................18
3.9.2 碳/氮源濃度的影響..........................18
3.9.3 培養體積...................................18
3.9.4 培養溫度...................................18
3.9.5 較適培養時間...............................19
3.10 微生物生長之分析............................19
3.11 表面張力測量................................19
第四章 結果與討論................................20
4.1 培養條件.....................................20
4.1.1 碳/氮源之選擇..............................20
4.1.2 各種水產廢棄物作為培養基對於靈菌紅素產量的影響...20
4.2 靈菌紅素之純化分離...........................22
4.3 以烏賊軟骨粉末作為培養基探討其他培養條件.....26
4.3.1 pH值之影響.................................26
4.3.2 碳氮源濃度之影響...........................28
4.3.3培養體積之影響..............................30
4.3.4 溫度之影響.................................31
4.4 靈菌紅素在不同pH值下的吸光值.................33
4.5 靈菌紅素在菌體內與上清液的比例比較...........34
4.6 照光與避光培養對於生產靈菌紅素的差異性.......35
4.7 靈菌紅素與表面張力值.........................37
4.8 其他不同碳氮源作為培養基的表面張力值.........38
4.9 其他不同碳氮源作為培養基的靈菌紅素生產量.....38
第五章 結論......................................42
參考文獻.........................................43

圖目錄
頁次
圖3.1. 實驗架構..................................13
圖4.1 TKU011於不同碳氮源培養基生產靈菌紅素之變化情形...21
圖4.2 prodigiosin之1H-NMR 的圖譜.................23
圖4.3 prodigiosin之13C-NMR 的圖譜................24
圖4.4 prodigiosin之ESI-MASS 圖譜.................25
圖4.5 TKU011不同pH值下與添加碳酸鈣後.............27
圖4.6 TKU011不同碳源濃度培養基與添加NPB緩衝液後..29
圖4.7 TKU011不同培養液體積生產靈菌紅素之變化情形.30
圖4.8 TKU011不同溫度培養基之
（A）菌體生長（B）生產靈菌紅素之變化情形.........32
圖4.9以OD535測量靈菌紅素在不同的pH值緩衝液之吸光值....33
圖4.10  上清液與沉澱物的靈菌紅素濃度比較.........34
圖4.11 照光與避光對於靈菌紅素生產的變化..........36
圖4.12 表面張力值與靈菌紅素生產濃度的比較........37
圖4.13 TKU011在不同培養基所得之
（A）表面張力值、（B）靈菌紅素濃度...............39

表目錄
頁次
表4.1 TKU011生產靈菌紅素之較適培養條件...........40
表4.2 不同文獻所載的prodigiosin類似物產量比較....41

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