The Effect of Upstream Performance of Biocatalytic Reaction on Downstream Purification Feasibility in Glycine Production Using Immobilized Aminoacylase

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羅思德(Rosyid Shidiq Hidayatulloh) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

(The Effect of Upstream Performance of Biocatalytic Reaction on Downstream Purification Feasibility in Glycine Production Using Immobilized Aminoacylase)

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摘要(中)

本研究探討使用活性碳上的固定化氨基化醯化?持續生產與純化甘氨酸，並將重點放在上游與下游製程。在上游階段，酵素反應顯示出有效的甘氨酸產量，固定化酵素展現出顯著的操作穩定性與可重複使用性。雖然酵素活性在第二週期下降至 18.2%，在第三週期下降至 9.5%，足以支持重複反應，驗證了酵素固定化長期使用的實用性。利用甘氨酸的三元相圖來優化下游的純化過程，從而了解甘氨酸結晶的最佳條件。在前兩個循環中，反應轉換率均在相圖的有利區域內，因此成功地純化了高純度的甘氨酸，純度範圍從 95.96% 到 102.6%，證實了純化方法的有效性。甘氨酸的產率介於 1.34% 到 5.67% 之間，與理論預測相當接近。然而，在第三個循環中，由於反應條件的轉變，純化不可行，落在了平衡區之外。這些發現突顯了優化上游生產和下游純化以提高產量和純度的重要性。本研究有助於發展甘氨酸生產的永續與綠色化學方法。

摘要(英)

This study investigates the sustainable production and purification of glycine using immobilized aminoacylase on activated carbon, focusing on both upstream and downstream processes. In the upstream phase, the enzymatic reaction demonstrated effective glycine production, with immobilized enzymes exhibiting significant operational stability and reusability. Although enzyme activity decreased into 18.2% in the second cycle and 9.5% in the third cycle, it remained sufficient to support repeated reactions, validating the practicality of enzyme immobilization for long-term use. The downstream purification process was optimized using a quasi-ternary phase diagram of glycine, which provided insights into the optimal conditions for glycine crystallization. Successful purification was achieved in the first two cycles, where reaction conversions fell within the favorable regions of the phase diagram, resulting in high-purity glycine with purity levels ranging from 95.96% to 102.6%, confirming the efficacy of the purification method. The yield of glycine varied between 1.34% and 5.67%, which is close enough with theoretical predictions. However, purification was not feasible in the third cycle due to a shift in reaction conditions, falling outside the equilibrium zone. These findings highlight the importance of optimizing both upstream production and downstream purification for improving yield and purity. This study contributes to the development of sustainable and green chemistry methodologies for glycine production.

關鍵字(中)

★ One keyword per line

關鍵字(英)

★ Enzymatic reaction
★ Crystallization
★ Purification
★ Phase diagram
★ Enzyme immobilization

論文目次

摘要... i
Abstract... ii
Acknowledgment... iii
Table of Contents... v
Table of Figures... viii
List of Tables... xi
Appendix... xii
Chapter 1. Introduction... 1
1.1 Background of the Study... 1
1.2 Upstream Process Overview... 6
1.3 Phase Diagram for Downstream Process... 8
1.4 Literature Survey on Enzyme Immobilization... 10
1.5 Conceptual Framework... 15
Chapter 2. Experimental Materials and Methods... 17
2.1 Materials and Chemicals... 17
2.2 Experimental Methods... 18
2.2.1 Solubility Measurements of Glycine, N-Acetylglycine and Sodium Acetate Trihydrate... 18
2.2.2 Enzymatic Reaction by Free Aminoacylase... 18
2.2.3 Immobilization of Aminoacylase... 19
2.2.4 Enzymatic Reaction by Immobilization Aminoacylase... 20
2.2.5 Conceptual Illustration of Quasi-Ternary Phase Diagram ... 21
2.2.6 Construction of Quasi-Ternary Phase Diagram... 25
2.2.7 Purification of Reacting Solution to Obtain Pure Glycine... 27
2.3 Analytical Measurements... 30
2.3.1 Fourier-Transform Infrared Spectroscopy (FTIR)... 30
2.3.2 Ultraviolet-Visible Spectroscopy (UV-Vis)... 30
2.3.3 Powder X-Ray Diffraction (PXRD)... 34
2.3.4 Scanning Electron Microscopy (SEM)... 35
2.3.5 Brunauer-Emmett-Teller Analysis (BET)... 35
Chapter 3. Results and Discussion... 36
3.1 Upstream Process for Glycine... 36
3.1.1 Immobilization of Aminoacylase on Activated Carbon... 36
3.1.2 Enzymatic Reaction with Free Enzyme and Immobilized Aminoacylase... 44
3.2 Downstream Process of Glycine... 55
3.2.1 Solubility Measurements of Glycine, N-acetylglycine, and Sodium Acetate Trihydrate... 55
3.2.2 Construction of Quasi-ternary Phase Diagram... 57
3.2.3 The Use of Quasi-Ternary Phase Diagram for Glycine Purification... 61
Chapter 4. Conclusions and Future Work... 68
4.1 Conclusions... 68
4.2 Future Work... 71
References... 72
Appendix... 79

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指導教授

李度(Tu Lee)

審核日期

2025-1-17

推文