本研究利用生物法產氫。Clostridium 是目前研究產氫的主要菌
株之一，本實驗所使用的Clostridium butyricum 在生長過程中，主要
的代謝酸有丁酸、醋酸，隨著丁、醋酸濃度的增加會有抑制菌體生長
的情形。本實驗欲利用常見的兩水相系統，探討以polyethylene
glycol(PEG)、Dextran(DEX)組成之兩水相系統，對於不同的物質有不
同的分配率的特徵，來減少或延緩代謝酸的抑制。
本實驗分四部分討論。第一部分是進行兩水相不同濃度組成中
丁、醋酸及菌體的分配情形；第二部分以血清瓶實驗觀察不同的兩水
相系統下，菌株實際的生長情形及產氫量；第三部分是利用2L 醱酵
槽進行批次醱酵；第四部分進行連續式饋料反應，進行饋料的依據一
般而言均是以碳源的使用情形來判斷是否進料，本實驗也是利用此原
則進行進料的動作。分配率實驗結果顯示，無論是丁酸、醋酸的分配
率皆大於1，且高分子濃度愈大，分配率也愈大，代謝酸都較偏好於
PEG-rich 相，而菌體則偏好於DEX-rich 相。適當的兩水相組成不會
抑制菌體的生長。比較有無兩水相系統的批次發酵實驗中，含有兩水
相系統的產氫量較無兩水相系統的產氫效率(Efficiency of H2
formation)約增加了18%。在連續式饋料(fed-batch)實驗中，含有兩水
相系統的產氫量與無兩水相系統的產氫效率則相近，其產氫效率68%
並無低於無兩水相系統的產氫效率67%，兩水相系統在整個醱酵(醱
酵時間36.5 小時)產氫停止前總共進料8 次，無兩水相系統(醱酵時間
36 小時)則進料4 次即停止產氫，含兩水相系統的連續式饋料實驗，
氫氣產率195.6 ml/hr/L，醱酵時間內產氫量共17136 ml。兩水相系統
有提昇Clostridium butyricum 產氫之現象。

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