| Summary: | 碩士 === 國立成功大學 === 化學工程學系 === 89 === This study is about producing creatinase by utilizing a recombinant strain, Escherichia coli JM109/pSCR05. The study focused on the effects of fermentation conditions on enzyme yield and plasmid stability. When cells grew in disadvantageous cultural environments, not only the growth rate decreased, but also the enzyme yield decreased. Several factors had been found to have influences on plasmid stability, including nutrient composition, pH, temperature, and oxygen transfer rate. In addition, the study explored the validity of the production of creatinase by fill-and-draw fermentation. Enzyme production was found to stop after the first medium exchange, suggesting that the recombinant strain could only be induced once by IPTG. According to this result, the amount of inducer required for the whole process should be added, all at once, during the batch phase.
中文摘要
英文摘要
目錄
表目錄
圖目錄
第一章 緒論
1-1 前言
1-2 肌酸酵素簡介
1-3 研究目的
第二章 實驗材料和方法
2-1 實驗材料
2-1-1 宿主-載體系統
2-1-2 藥品
2-1-3 實驗儀器與裝置
2-2 實驗方法
2-2-1 菌種保存
2-2-2 培養基
2-2-3 醱酵實驗
2-2-4 溶氧電極之校正方法
2-3 分析方法
2-3-1 菌體濃度檢量線製作方法
2-3-2 菌體濃度分析方法
2-3-3 酵素液之取得
2-3-4 酵素的活性分析
2-3-5 質體穩定性之測定
2-3-6 繼代培養
第三章 結果與討論
3-1 培養世代對質體穩定性之影響
3-2 搖瓶和醱酵槽培養對質體穩定性之影響
3-3 培養條件對酵素生產及質體穩定性之影響
3-3-1 氧氣傳送速率之影響
3-3-2 pH值之影響
3-3-3 溫度之影響
3-4 培養基組成對酵素生產及質體穩定性之影響
3-4-1 葡萄糖濃度之影響
3-4-2 酵母萃取物之影響
3-4-3 消化蛋白之影響
3-4-4 磷酸鹽之影響
3-5 以半連續式醱酵生產肌酸酵素
3-5-1 半連續式醱酵：每小時置換200 mL、不添加Ampicillin
3-5-2 半連續式醱酵：每3小時置換300 mL
3-5-3 半連續式醱酵：每6小時置換1000 mL
3-5-4 半連續式醱酵：每6小時置換1000 mL、控制溶氧值
3-5-5 半連續式醱酵：每6小時置換1000 mL、LB培養基
3-5-6 批次醱酵：在前培養中添加IPTG
3-5-7 半連續式醱酵：每3小時置換300 mL、於批次相一次添加所需之IPTG
第四章 結論
第五章 未來展望
參考文獻
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