臺灣博碩士論文加值系統

本研究成功地使用Novozym 435為觸媒，來合成咖啡酸苯乙基酯。本研究發現咖啡酸在非極性溶液裡的溶解度雖然很低，但其反應速率卻比在極性溶劑中快。本研究使用異辛烷作為反應溶劑、70℃為反應溫度，可增加酵素的活性與反應物的溶解度。苯乙醇的最佳反應量為125 mg，過量的醇會導致酵素失活。使用30 mg的酵素量能於兩天內使轉化率達到近100%。本研究亦發現磁石之攪拌轉速與轉化率並無太大關係，酵素在第三次重複使用後仍可達到原先90%之活性。在高轉速下，重複使用的酵素很快失去活性；若不使用磁石攪拌，酵素穩定性較佳，但轉化率低於90%。

Enzymatic synthesis of caffeic acid phenethyl ester from caffeic acid and 2-phenylethanol has been successfully carried out using Novozym 435 as the catalyst. Although the solubility of caffeic acid was low in non-polar solvents, they were found to give better conversions than in polar solvents. Isooctane gave the highest conversion and was chosen as the solvent in this study. A temperature of 70oC was proven as the optimum reaction temperature. This was attributed to the increase in enzyme activity and solubility of reactants. At a fixed caffeic amount of 2 mg, the optimum alcohol amount was 125 mg, higher alcohol amount resulted in the inactivation of enzyme. An enzyme load of 30 mg gave nearly 100% conversion in 48 h. The conversion was found to be independent of stirrer speed. The reuse stability of enzyme showed that enzyme could maintain more than 90% of its original activity up to the third batch. Enzyme lost its activity faster under higher stirrer speed. Reactions without stirring gave better enzyme stability, however the conversions were always less than 90%.

摘要.............................................................Ⅰ
Abstract.........................................................Ⅱ
致謝.............................................................Ⅲ
Contents.........................................................Ⅳ
Figure legends...................................................Ⅵ

Chapter 1 Introduction............................................1
1.1 CAPE..........................................................1
1.2 Antioxidants..................................................1
1.3 Radical scavenging............................................3
1.4 Mechanism of antioxidant......................................5
1.5 CAPE as a radical scavenger in a typical pathologic process...6
1.6 Enzymatic synthesis of CAPE...................................7
1.7 Motivation of this research...................................8

Chapter 2 Literature survey.......................................9

2.1 Chemical method to synthesize CAPE............................9
2.2 Helping the environment through the use of enzymes...........10
2.3 Enzymatic activity in organic solvents.......................11
2.4 Lyophilized and crystalline enzymes..........................12

Chapter 3 Materials and methods..................................14
3.1 Chemicals....................................................14
3.2 Equipments...................................................15
3.3 Synthesis of CAPE............................................15
3.4 Reuse stability of enzyme....................................16
3.5 HPLC analysis................................................17
3.6 Determination of caffeic acid conversion.....................17
3.7 Assay of enzyme activity.....................................19

Chapter 4 Results and discussions................................20
4.1 Selection of enzymes.........................................20
4.2 Effect of solvent (log P)....................................21
4.3 Effect of temperature........................................22
4.4 Effect of alcohol amount.....................................24
4.5 Effect of the amount of enzyme...............................27
4.6 Effect of stirring speed.....................................28
4.7 Reuse stability study of enzyme..............................28

Chapter 5 Conclusion.............................................31

Reference........................................................33
作者簡介.........................................................41

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