單加氧酶存在於不同的生命系統中，也存在人類肝臟的微粒體內，具有類固醇與脂肪酸羥化等能力，在生物降解以及代謝扮演著相當重要的角色。此系統中的酵素細胞色素P450 BM-3，源於巨大芽孢桿菌，具有氧化長碳鏈(C12-C16)脂肪酸羥基化的能力，催化活性中心是由含鐵紫質(heme)所構成，我們利用大腸桿菌來大量表現P450 BM3，並且利用此系統來催化反應受質，同時合成出不同類型的含氟探針，來探討細胞色素P450 BM3對於不同受質的變異性。並使用氣相層析質譜儀等儀器來分析氧化後的產物，就反應的結果來看，我們發現在末端氟原子的數量會影響反應的活性以及產物的氧化位置，對比晶體文獻的結果，推測在Cytochrome P450 BM3的疏水性口袋中，末端氟原子與口袋內部活性中心附近的胺基酸有著微小力在作用，末端氟原子數量的上升會影響受質在活性口袋反應的位置，藉此影響受質與活性中心反應的專一性。
脂肪水解酶(lipase B)能夠在水溶液中轉化三酸甘油酯水解成甘油跟脂肪酸，而有機溶劑條件下則可促進酯類合成甚至轉酯化，存在酵母菌中的脂肪水解酶CALB, 有機溶劑中能夠對於中碳鏈長度的烷醇進行選擇性酯化，經由固定於丙烯酸樹酯的脂肪水解酶，不只是在二號醇位置有選擇性，針對三號醇類化合物也會有光學選擇性，因此我們利用此酵素催化不同光學性質的3-辛醇使用氣相層析儀比較產物的差異性，搭配使用(R)-(-)-O-乙醯基扁桃酸的衍生化反應後利用核磁共振光譜交叉證實，在三號醇位置在(R)-3-辛醇可以酯化，在證實了辛烷在二號醇與三號醇可以選擇R-form的前提下，將脂肪酶與P450 BM3的產物作用，利用脂肪酶來鑑別被反應受質的光學性質，最後成功的發現在1-氟辛烷與Cytochrome P450 BM3作用的產物能夠接續脂肪酶酯化反應，而得到含氟產物被CALB選擇的證據。

Oxidizing C–H bond of organic compounds is an issue in organic chemistry. But some enzymes have the ability to oxidize organic compound. The enzyme called Cytochrome P450 BM3 from Bacillus megaterium which is able to oxidation of long chain fatty acid (C12-C20). We are interesting for what happen in the P450 BM3 reaction pocket, so we designed the fluorinated probe to observe the reaction.

On the other hand, we found another enzyme called Candida antarctica Lipase B (CALB) which has ability to selec transesterification for (R)-2-heptanol and (R)-2-pentanol , and we successfully proof transesterification in 2-pentaol to 2-dodecanol ,which has good enantiomeric excess (ee), then we changed different OH site and found that 3-octanol also has the same property. We want to use this assay combine with the P450 BM3 hydroxylation to know the enantiomeric excess on w-2,w-3 compound P450 BM3 catalyzed. Finally we successfully found the case when 1-fluorooctane is oxidation by P450 BM3, the w-2 site is major R-form formation and w-1 is major in S-form formation.

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