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中國畜牧學會會誌45(增刊) |
,2016 |
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J. Chin. Soc. Anim. Sci. 45 (Suppl.) |
147
69.自活性污泥篩選具脂解活性之細菌
廖仁寶 陳若菁 蕭庭訓 程梅萍 吳明哲
行政院農業委員會畜產試驗所
本研究目的為自畜產廢水處理設施中之活性污泥樣品，篩選能產生胞外脂解酵素的細菌，期 能應用於生物產業。以傳統菌株分離篩選方式，將活性污泥樣品溶於無菌水中，序列稀釋後 塗抹於選擇性培養基上，分別在 37°C 與 55°C 下過夜培養。在選擇性培養基上可發現一些 藍色菌落與其周圍有透明圈，此為能產生胞外脂解酵素之菌株，進一步將此等菌株純化培養 與冷凍保存。總共篩選出 9 株具脂解活性菌株，其中於 37°C 培養條件下篩得 5 株；在 55°C 培養條件下則篩得 4 株。進一步培養這 9 株菌，並萃取其 DNA 以進行分子鑑定。由 16S rRNA 基因序列分析得知，此 9 株菌中，5 株為 Bacillus licheniformis、2 株為 Bacillus pumilus、1 株為 Bacillus subtilis 及另 1 株為 Bacillus mojavenis。其中 B. mojavenis 為較特 殊的菌株，將列為進一步研究的對象。
關鍵語：活性污泥、脂解活性、選擇性培養基
Isolation of lipolytic enzyme producing bacteria from activated sludge
R. B. Liaw, J. C. Chen, T. H. Hsiao, M. P. Cheng and M. C. Wu
Livestock Research Institute, Council of Agriculture, Executive Yuan The purpose of this study was to screen for lipolytic enzyme producing bacteria from activated
sludge samples of livestock wastewater treatment facility, and these isolates were expected for
application in bioindustry. By conventional isolating protocol, the activated sludge samples were
diluted in series in sterile water and smeared on selective media. The media were grown at 37°C
and 55°C, respectively, overnight. Some isolates showed lipolytic activities with clear zones around
their colonies on the selective media, and these isolates were purified and preserved in a freezer. A
total of nine isolates were obtained, 5 isolates cultured at 37°C and 4 isolates cultured at 55°C.
Furthermore, the nine isolates were cultured for DNA extraction and molecular identification.
Based on the analysis of 16S rRNA gene sequences, 5 isolates were identified as Bacillus
licheniformis, 2 isolates as Bacillus pumilus, others as Bacillus subtilis and Bacillus mojavenis,
respectively. Because B. mojavenis was a special one among these isolates, it was the first target for
future study.
Key Words: Activated sludge, Lipolytic activity, Selective medium