Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3071
標題: 利用蕈狀芽孢桿菌與枯草桿菌生產生物界面活性劑之液態培養基探討
Production of biosurfactant by Bacillus mycoides NP02 and Bacillus subtilis CWS1 in submerged culture
作者: 周昀蓉
Chou, Yun-Jung
關鍵字: 蕈狀芽孢桿菌
Bacillus mycoides
枯草桿菌
生物界面活性劑
油脂添加
最佳化
Bacillus subtilis
biosurfactant
oil addition
response surface methodology
出版社: 化學工程學系所
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摘要: 近年來,生物防治已成為農業發展之目標,即是利用自然界所存在的拮抗微生物減緩病害發生速率,並達到病害防治的效果。本研究目的以具有抑制植物病原菌之蕈狀芽孢桿菌Bacillus mycoides NP02液態醱酵生產生物界面活性劑。研究結果於不同組成培養基進行液態醱酵培養,以peptone為培養基之過程中可得較高的生物界面活性劑產量為71.27±1.93 mg/l。 另外針對不同添加物至培養基中對生物界面活性劑產量影響,發現玉米油的添加可使醱酵液的表面張力降低,其中以peptone為培養基並添加玉米油可使醱酵液之表面張力降到33 mN/m,以HPLC分析發現其生物界面活性劑產量高達1744.81±144.89 mg/l,與未添加油脂前之產量相差25倍,證實油脂的添加可增加生物界面活性劑產量。以HPLC分析由Bacillus mycoides NP02所產出的生物界面活性劑,其分析圖譜於14.6 min發現不同於surfactin標準品之成分波峰,但仍具有降低表面張力之活性,此生物界面活性劑可推測極為可能為surfactin isoform,因此需要進一步去鑑定。 以枯草桿菌Bacillus subtilis CWS1生產生物界面活性劑,在不同碳源試驗中,以sucrose為最佳碳源,其生物界面活性劑產量為2283.09±86.85 mg/l;在不同氮源測試中,以(NH4)2SO4為最佳氮源,其生物界面活性劑為2613.16±49.69 mg/l。在培養基最佳化實驗中,選擇碳源、氮源以及玉米油做為設計因子以反應曲面法找出最佳之培養基組成,結果顯示sucrose 50 g/l、(NH4)2SO4 3 g/l以及玉米油 5 g/l於30℃之操作條件進行液態培養,預測可得最佳的生物界面活性劑濃度為8057.57 mg/l。
In recent years, microbial pesticides for biocontrol control has become the goal of agricultural development. In this study, the production of biosurfactant by Bacillus mycoides NP02 under peptone-based medium was found to reached 71.27�1.93 mg/l. Various types of additives were added into main culture. It was observed that corn oil could reduce the broth tension to 33 mN/m with peptone-based medium, and its biosurfactant production could reach 1744.81�144.89 mg/l, which was approximately 20-fold to that of the control. This result demonstrates oil addition to medium in submerged culture could enhance the biosurfactant production. The biosurfactant produced by B. mycoides NP02 was analyzed with HPLC, and its retention time of 14.6 min is different from that of the surfactin standard. This unknown biosurfactant in broth is expected to be a surfactin isoform, which need further identification. On the other hand, the optimizing medium enhanced biosurfactant production by Bacillus subtilis CWS1. In carbon source study, sucrose gave the best biosurfactant production, reaching 2283.09�86.85 mg/l. In nitrogen source study, ammonium sulfate was identified to be the most favorable source for biosurfactant production, reaching 2613.16�49.69 mg/l. In the medium optimization, carbon source, nitrogen source and corn oil were chosen as designing factor. According to the response surface methodology, the optimal concentrations of sucrose, ammonium sulfate and corn oil were 50 g/l, 3 g/l and 5 g/l, respectively, where the best biosurfactin production reached 8057.57 mg/l.
URI: http://hdl.handle.net/11455/3071
其他識別: U0005-1708201211410900
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1708201211410900
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