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研究生:王秋玲
研究生(外文):Chung Lang Wang
論文名稱:利用Pseudomonasaeruginosa生產醣脂類界面活性劑
論文名稱(外文):Production of rhamnolipid biosurfactants by Pseudomonas aeruginosa
指導教授:徐泰浩徐泰浩引用關係
指導教授(外文):Tai-Hao Hsu
學位類別:碩士
校院名稱:大葉大學
系所名稱:食品工程研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:1995
畢業學年度:83
語文別:中文
論文頁數:119
中文關鍵詞:綠膿桿菌生物界面活性劑鼠李糖脂
外文關鍵詞:Pseudomonas aeruginosaBiosurfactantsRhamnolipid
相關次數:
  • 被引用被引用:2
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本研究以綠膿桿菌( Pseudomonas aeruginosa CCRC11633 )與螢光假
單胞菌(Pseudomonas fluorescens CCRC14347 )生產生物界面活性劑。
在多次重覆試驗中,發現在相同培養條件下以 Pseudomonas aeruginosa
所分泌之界面活性物質產量較高。經由液體營養培養基與五種不同碳源(
葡萄糖、蔗糖、橄欖油、大豆油、石蠟)等六種不同的培養基培養之結果
顯示,綠膿桿菌以橄欖油和大豆油為碳源培養時,可使發酵液之表面張力
可降至 35 dyne/cm 以下。 而後,再以甘油 - 蛋白月東培養基培養,其
發酵液之表面張力降低至 26.2 dyne/cm。 以石碳酸 - 硫酸法分析含糖
之產物,再經薄層層析鑑定含有鼠李糖,其產量可達 6.47 g/L。若利用
二階培養可使產量高達 13.21g/L。 在搖瓶試驗時,最適之培養溫度、酸
鹼值、培養體積、碳與氮源,分別為 30 ℃、pH7.0、每升300mL 培養基
、5%甘油與4%蛋白棟。 此物質以薄層層析法鑑定含有鼠李糖,臨界微胞
濃度為 1.1mL/L,在高溫、高壓下、在不同 pH 下具有穩定性。在回收方
法上以超過濾法較溶劑萃取方法佳。鼠李糖脂利用陰離子交換樹脂(
DEAE-Sephacel )溶離在 0.8M NaCl Tris-HCl(pH8.0) 緩衝溶液中, 可
將之純化出,其表面張力為 26.7 dyne/cm。 就生理而言,具有溶血與抑
菌作用,結果顯示 10 μ g鼠李糖脂對於 Escherichia coli、
Bacillus cereus、Bacillus subtilis 與Staphylcoccus aureus 等四株
菌皆有抑菌作用,隨著濃度增加對E. coli 和Staphylcoccus aureus 之
抑菌效果愈大。採集彰化和嘉義受油污染地區之土壤,利用含油之培養基
進行菌種篩選,發酵液之表面張力可降至 30.1 dyne/cm。
In this study,Pseudomonas aeruginosa CCRC11633 and Pseudomonas
fluorescens CCRC14347 produced biosurfactants. The result of
showed that P.aeruginosa had the highest level of biosurfactant
secretion among tested strains. Nutrient broth and five
different carbon sources(glucose、surcose、olive oil、 soybean
oil、n-paraffin )media were tested on shaker-flask scale for
culturing P. aeruginosa with olive oil and soybean oil as
carbon source,the surface tension of P. aeruginosa
fermentation broth were red uced to 35.0 dyne/cm and the
surface tension of fermemtation broth were reduced to 26.2 dyne/
cm. When glycerol- peptone were applied in the medium. We used
spectrophotometer to monitor the secretion of rhamnose of the
P. aeruginosa,the yield were 6.47 g/L. Use two-stage culture,
the yield were 13.21g/L. The optiumn cultural condition: with
fermentation temperature of 30 ℃、pH7.0、300mL/L of medium、5%
Glycerol as carbon and 4% peptone as nitrogen source. The
biosurfactants produced by P. aeruginosa were identified by TLC
as rhamnose,the CMC is 0.11 mL/dL and were stable at high
temperature(121 ℃ for 20mins)and different pH value
ranges(1-14). Ultrafiltration rather than extraction were
better in recovery of rhamnolipid. Anion exchange
chromatography on DEAE-Sephacel was chosen for further purifi-
cation. The rhamnolipid was eluted from the matrix with 0.8M
NaCl Tris-HCl buffer and the surface tension of this elution
was 26.7 dyne/cm. Ten μ l of rhamnolipid showed growth
inhibition to Escherichia. coli、Bacillus cereus、Bacillus
subtilis and Staphylcoccus aureus. Comply with rhamnolipid
concertration increase, the inhibition ring augment for E.
coli and Staphylcoccus aureus. Using oil-containg medium,
biosurfactant producing strain with low surface tension(30.1
dyne/cm)was isolated from Chang-Hwa and Jia-yi area.
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