以脂肪酶固定化金屬有機骨架材料進行生物性催化合成華法林抗凝血劑

Lipase Immobilized Metal-Organic Framework as a Biocatalyst for Warfarin Synthesis

楊妮欣(Ni-Shin Yang)

指導教授 ： 黃悉雅

中原大學/理學院/化學研究所/碩士(2013年)

https://doi.org/10.6840/cycu201301002

摘要

有鑑於生物催化備受工業與學術界的重視，但傳統溶液態的催化方式，具有酵素無法回收、催化壽命短以及熱不穩性等缺點，因此本論文開發新穎性酵素固定化微反應器，以克服溶液態之缺點。
研究中利用物理吸附方式，將脂肪酶固定於金屬有機骨架材料形成脂肪酶固定化微反應器，並進行生物催化合成華法林抗凝血藥物。首先建立微胞電層析方法，監測其反應物4-羥基香豆素和亞芐基丙酮與催化產物華法林的變化情形，以進行產率效能評估。其次利用殼聚醣塗覆之毛細管，並以γ-環糊精作為對掌選擇器之毛細管電泳方法，分離華法林產物之對掌異構物。
結果顯示，以UiO-66(Zr)金屬有機骨架材料所製備之脂肪酶固定化微反應器，可在反應一天後催化生成華法林，並重複使用五次後，產率依然可達70%以上，優於傳統溶液態催化合成方式(61%, 只能催化一次)。研究中再進一步測試以不同種類之金屬有機骨架材料固定脂肪酶，並催化合成華法林，其中以UiO-66(Zr)催化產率最高(第一次產率達86.9%)，其原因推測是UiO-66(Zr)較UiO-66-NH2(Zr)具有疏水作用力，利於酵素吸附。最後與常見之中孔矽材SBA-15相互比較，結果顯示，本研究所開發之酵素微反應器的催化產率更佳。因此，本實驗所開發的酵素固定化微反應器，具有重複使用、催化時間短、易純化產物等優點，於華法林的催化反應上是相當適合的。

關鍵字

金屬有機骨架 ； 華法林 ； 脂肪酶固定化

並列摘要

Biocatalysis has attracted more attention in industry and academia fields. There are some disadvantages such as difficult recycling, short catalytic lifetime and thermal unstability in traditional in-solution catalysis. In this study, novel enzyme immobilized microreactors were developed to overcome the disadvantages occurred in in-solution method.
Herein, a lipase immobilized microreactor was performd by lipase enzyme physically adsorbed onto metal-organic frameworks (MOFs) (named as lipase@MOFs) and the lipase@MOFs were used as biocatalyst for anticoagulants warfarin synthesis. First, a micellar electrokinetic chromatography (MEKC) method was established to quantify the reactants (4-hydroxycoumarin and benzylideneacetone) and the product (warfarin) for catalytic yield evaluation. Next, using chitosan coated column to analyze enantiomer of warfarin products by capillary electrophoresis with γ-cyclodextrin (γ-CD) as a chiral selector.
Results indicated that the warfarin yield produced the lipase immobilized UiO-66(Zr) microreactor under 50°C and 1 day for 5th reuses was still above 70%, which is better than traditional lipase in-solution catalysis (61% for only single use). Furthermore, various MOFs were used to immobilize lipase for warfarin synthesis, among them UiO-66(Zr) as immobilized microreactors for warfarin synthesis has the best warfarin yield (up to 86.9%, 1st reuse). The reason for that was possibly because due to the increased hydrophobicity is helpful for enzyme immobilization. Finally, compared with the common mesoporous silica materials, SBA-15, this study indicated that MOFs can provide a better catalytical ability. Consequently, a suitable lipase immobilized microreactor for warfarin synthesis that provided advantages such as reusability, catalytic short time and easily purification was developed.

並列關鍵字

catalysis ； immobilized ； lipase ； MOF

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