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研究生: 陳秀娟
Chen Hsiu-Chuan
論文名稱: 污水中分離外分泌性脂質酵素細菌的鑑定與其生理特性之研究
Identification and physiological characterization of extracellular lipase producing bacteria isolated from the sewage
指導教授: 李銘亮
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
畢業學年度: 86
語文別: 中文
中文關鍵詞: 脂質酵素
論文種類: 學術論文
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  • 從污水中篩選出來的細菌三株 I3a, II11 和 II11' 用 Mg medium 培養,此培養基含有牛油作為碳的唯一來源. 此三菌株 I3a, II11 和 II11' 經過鑑定,分別為 Pseudomonasaeruginosa, Aeromonas hydrophila 和 Acinetobacter baumanii. 此三菌株在光學顯微鏡下觀察均為桿狀,且大小都比大腸桿菌小,因此,用電子顯微鏡觀察更詳細的外型. 第一菌株綠膿桿菌(Pseudomonas aeruginosa I3a)在 30C 以及 pH7.0 產生脂質酵素的量最高,而且 30C 也是生長最佳溫度. 以各種 p-nitropheny esters 當受質, 此脂質酵素對 C12 醯基感受性最高. 同時脂質酵素活性會受汞離子的抑制, 而Mn2+, Mg2+ 和 Ca2+ 對此脂質酵素活性並無影響. 氣單胞菌(Aeromonas hydrophila II11)在 30C 產生脂質酵素的總量最好,也是最佳的生 長溫度,同時在 pH6-pH8 的環境產生脂質酵素較高. 此脂質酵素對於醯基並沒有專一性. 此脂質酵素活性會受Hg2+ 抑制, Mn2+ 和 Ca2+ 沒有很明顯的影響, 但 Mg2+ 對此脂質酵素活性卻有促進作用. 第三菌株靜止桿菌(Acineto. baumanii II11')是在 37C 表現脂質酵素量最高,最適合生 長溫度為 30C ,而其表現的最適合 pH 值為 pH8. 脂質酵素對 C14 醯基反應較靈敏. 同時, 其脂質酵素活性受 Hg2+, Mg2+ 和 Mn2+ 抑制, Ca2+ 對此脂質酵素活性並無影響.上述三菌株的分類, P. aeruiginosa I3a 是屬於需氧菌,A. hydrophila II11 為兼性需氧菌, acineto. baumanii II11' 是需氧菌. 共同特性是脂質酵素均屬於鹼性脂質酵素.但是產生脂質酵素的最適 pH 值各不相同, 而脂質酵素對溫度的耐受性也不一樣.

    Three bacteria strains (I3a, II11 and II11') were isolated from the sewage by using a Mg medium which contained butter as the sole carbon source. Thesethree strains were identified as namely; Pseudomonas aeruginosa, Aeromonashydrophila and Acinetobacter baumanii. They were observed under the dark fieldmicroscope as morphologially rod-sheped, and their sizes examined were smallerthan E. coli. So, they were further identified and examined in more detail with electron microscope. The highest amount of lipase produced by P. aeruginosa I3a is at 30C, and itsoptimal growth pH is around 7. With p-nitrophenyl esters as its substrates, theenzyme shows a preference for C12 acy group. The enzyme activity was inhibited byHg2+ and Mn2+, but did not significantly affect by the Mg2+ and Ca2+. The highest amount of lipase produced by A. hydrophila II11 is at 30C, and they produced lipase better at pH6-pH8, this enzyme shows no preference for anysubstrate. The enzyme activity was inhibited by Hg2+, and not affect by Ca2+ andMn卻有促進作用. The highest amount of lipse produced by Acineto. baumanii II11' is at 37C, but30C is the optimal growth temperature. Its optimal pH of lipase producing is 8. The enzyme shows a preference for C4 acy group. The enzyme activity was inhibitedby Hg2+, Mg2+, and Mn2+, but did not significantly affect by the Ca2+. In classification, P. aeruginosa I3a is aerobes, A. hydrophila II11 is facutauiveanaerobes, and Acineto. baumanii II11' is aerobes. They possess alkaline lipases,however their optimal pH of lipase production are different. Besides, sensitivityof three enzyme to various temperatures are different.

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