利用嗜鹼性芽胞桿菌Bacillus circulans No. 38-2 (BCRC 10094)進行醱酵生產環狀糊精葡萄糖苷轉移酶之研究

Production of cyclodextrin glucanotransferase (CGTase) by Bacillus circulans No. 38-2 (BCRC 10094)

林清安(Ching-An Lin)

指導教授 ： 段國仁

大同大學/工程學院/生物工程學系所/碩士(2005年)

摘要

本研究利用嗜鹼性芽胞桿菌（Bacillus circulans No. 38-2，BCRC 10094）進行醱酵生產CGTase的研究。分別使用玉米澱粉和可溶性澱粉為碳源，能夠有效的誘導生產CGTase。而澱粉同時扮演碳源與誘導劑的功能。在33 C下，以基礎培養基與澱粉20 （g/L）為碳源的批式醱酵能生產約17319 Unit/ml的酵素活性。在pH-stat的饋料批式醱酵培養中，以澱粉的培養基，能夠得到約63165 Unit/ml的酵素活性。若先以葡萄糖為碳源將細胞的濃度提高，然後再饋入澱粉來誘導酵素，則能得到約39500 Unit/ml的酵素活性。在連續式醱酵培養的操作條件下，以稀釋率0.15倍進行CGTase的生產，結果能得到 31722 Unit/ml的酵素活性，以稀釋率0.3倍進行 CGTase的生產，則能得到約14320 Unit/ml的酵素活性。本結果顯示，利用pH-stat的饋料批式醱酵培養，在以澱粉為碳源的條件下，能夠得到最大量的酵素活性。若以單位體積產率（volumetric productivity）看則以連續式醱酵在稀釋率為0.3倍時，得到的酵素活性最高（約2864 Unit/h/L），其次為連續式醱酵在稀釋率為0.15倍（約2379 Unit/h/L），以pH-stat 饋料批式醱酵及批式醱酵以澱粉為碳源的體積生產速率分別為 （638, 444 Unit/h/L）。

關鍵字

嗜鹼性芽胞桿菌、環狀糊精葡萄糖苷轉移酶

並列摘要

Production of cyclodextrin glucanotransferase (CGTase) by Bacillus circulans No. 38-2 (BCRC 10094) in alkaline medium under various conditions was investigated. Corn starch and soluble starch were employed as the carbon source for culturing the strain and performing as an inducer of CGTase, The basal medium (2 % yeast extract, 1 % Na2CO3, 0.1 % KH2PO4, 0.1 % K2HPO4, 0.02 % CaCl2, 0.02 % MgSO4, 0.02 % Mn(NO3)2 ) and 2 % starch as carbon source could produce the CGTase activity of 17319 unit/ml in batch culture at 33 C. The CGTase activity could be achieved 63165 unit/ml using soluble starch as carbon source in pH-state fed-batch fermentation. Another fermentation strategy was performed by culturing the cell to high density using glucose as carbon source in pH-stat strategy, and then CGTase was induced by soluble starch to achieve 39500 unit/ml activity. In continuous culture, 31722 unit/ml and 14320 unit/ml were achieved at respective diluting rate of 0.15 and 0.3. The results indicated that the most volumetric activity (2864 unit/h/L) was achieved for continuous fermentation at a dilution rate of 0.3, and then at a dilution rate of 0.15 (2379 unit/h/L). The volumetric activities of 638 and 444 unit/h/L were achieved for fed-batch and batch fermentation respectively.

並列關鍵字

Bacillus circulans ； cyclodextrin glucanotransferase

參考文獻

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