探討解酯酶活性的研究已發展超過一世紀，其偵測方法有許多種，如滴定法、Rhodamine B agar plate…等，但使用現行方法偵測酶活較耗時且成本高，因此本研究嘗試建立快速響應之光學與電化學微系統來偵測Candida rugosa解酯酶活性。
光學微系統選用能穩定生成固定大小之靜止微液滴，探討酸鹼值與界面活性劑對解酯酶的影響，液滴內之游離酵素含有解酯酶與甘油螢光試劑，當系統酸鹼值為pH 7.0、pH 7.4、pH 8.6時，解酯酶相對活性為100 %、95 %、4 %；當系統甘油濃度為0.01 M且酸鹼值為pH 7.0、pH 7.4、pH 8.6時，甘油螢光試劑中的glycerol enzyme mix與glycerol probe相對活性為84 %、90 %、100 %。為了降低pH值對偵測靈敏度的影響，本研究最終選用電化學生物感測之微系統，以電極表面與電解質中化學反應造成的電荷變化來探討解酯酶活性。
電化學生物感測之微系統一開始利用交流阻抗法檢測甘油濃度來確定解酯酶活性。實驗結果顯示在緩衝液為HEPES時，電子轉移阻抗與溶液甘油含量呈線性關係(R2 = 0.94)，但因靈敏度低且標準差過大，改為將甘油去氫酶與NADH輔酶固定化於自組裝電極，並使用循環伏安法來檢測低於1 %之甘油溶液，結果顯示反應電流的大小在NADH氧化還原電位 - 0.31 V時與甘油濃度具相當高的線性關係（R2 = 0.99），但因響應值過低，最後再試圖利用長鏈自組裝膜改質電極，在循環伏安進行解酯酶活性測試，得到初始水解速率為2.42 ×〖10〗^(-5) M/s和平衡水解速率0.2 ×〖10〗^(-5) M/s。

The investigation of lipase has been developed for nearly a century. However, conventional methods for measuring activity of lipase are time-consuming and expensive. Therefore, the aim of this study is to establish a rapid response micro-system to investigate the lipase activity from Candida rugosa based on optical and electrochemical detections. The optical detection was used for studying the effects of pH and surfactant on the lipase activity. When the pH values were 7.0, 7.4 and 8.6, the relative lipase activity were 100 %, 95 %, and 4 %, respectively, while the fluorogenic enzyme activities were 84 %, 90 %, and 100 %, respectively. Because the enzymes applied in the glycerol fluorogenic kit was pH sensitive, it was disadvantage to use optical detection for future applications. Therefore, we further performed quantification of glycerol based on alternative current impedance; the results indicated that the standard deviation was higher than the sensitivity of the detection. Therefore, glycerol dehydrogenase and NADH were immobilized with cross-linking agents by forming a self-assembled monolayer (SAM) on a gold electrode. The SAM system could detect glycerol solution and lipase activity by the cyclic voltammetry. The results show that the initial and equilibrium hydrolysis rate of lipids by lipase was 2.42 ×〖10〗^(-5) M/s and 0.2 ×〖10〗^(-5) M/s, respectively. The detection sensitivity was as high as 0.62 vol% glycerol / μΑ These results provide the basis for future development of the rapid and affordable analysis technique for the lipase activity.

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