碳六十固定化脂肪?酯類光學異構物壓電晶體感測器研製與應用

陳秋樺

指導教授 ： 施正雄

國立臺灣師範大學/理學院/化學系/碩士(2002年)

摘要

本研究合成碳六十(C60)－固定化酵素：碳六十/脂肪分解酵素，並研製一酯類光學異構物壓電晶體感測器。將塗佈碳六十的石英壓電晶體感測器做為研究碳六十與脂肪分解酵素之間作用力的工具，結果顯示酵素能被碳六十所吸附並呈現不可逆的化學吸附現象。
探討塗佈碳六十的石英壓電感測器與脂肪分解酵素之間的pH及溶劑效應。碳六十與脂肪分解酵素作用的最適pH為7，最適溫度為31oC，在有機溶劑(甲醇、乙醇)存在時，碳六十與脂肪分解酵素的結合能力會降低。
將碳六十與酵素反應生成不溶於水的碳六十/脂肪分解酵素，並以IR鑑定產物，將產物譜圖與碳六十及酵素的譜圖比較後，能夠得知產物是由碳六十及酵素鍵結而成。以孔徑大小約5~10μm的多孔性二氧化矽材質的濾片浸泡於碳六十/甲苯溶液中，帶溶液揮發後得到碳六十濾片，將碳六十濾片浸泡於酵素(脂肪分解酵素)溶液中，製作碳六十/脂肪分解酵素，經測試後，發現所製作的碳六十/脂肪分解酵素濾片具有活性。當浸泡酵素溶液的濃度增加時，碳六十/脂肪分解酵素的催化能力也隨之增加。自行製作的碳六十/脂肪分解酵素濾片可以催化L－form胺基酸酯類水解產生L－form胺基酸，而對於D－form的胺基酸酯類則無水解反應發生。
使用塗佈C60－cryptand[2,2]的石英壓電感測器以及固定化的碳六十/脂肪分解酵素來偵測水溶液中的L-form胺基酸酯類。利用L-form胺基酸(L-form胺基酸酯類經碳六十/脂肪分解酵素水解後的產物)可被塗佈C60－cryptand[2,2]的石英晶體吸附的原理，使用自製的碳六十/脂肪分解酵素濾片研製一酯類光學異構物壓電晶體感測器。所研製的酯類光學異構物壓電晶體測器能成功的分辨D－form與L－form的胺基酸酯類，且對於L－form的胺基酸酯類的偵測具有良好的靈敏度及再現性。

關鍵字

碳六十 ； 脂肪分解酵素 ； 壓電感測器 ； 光學異構物

並列摘要

Immobilized fullerene C60－lipase was synthesized and applied in a piezoelectric crystal sensor for optical isomer of esters. The C60 coated piezoelectric quartz crystal sensor was employed to study the interaction between C60 and lipase. The partially irreversible response for lipase was obserred by the desorption study, which implied that lipase could be adsorbed on C60 by chemisorption.
Effects of pH and solvents on the response of the C60 coated piezoelectric crystal sensor for lipase was also studied. Optimum pH was found to be at 7 and optimum temperature was 31oC. The reactivity between C60 and lipase was decreased in organic solvents such as methanol and ethanol.
The water insoluble C60－lipase was identified with IR spectrography. The C60－lipase coated SiO2 chip with pore size of about 5~10 μm was prepared. The activity of immobilized C60－lipase was investigated and was still activated. The increased concentration of lipase resulted the increase in the ability of C60－lipase catalyze the hydrolysis of L－amino acid esters. The immobilized C60－lipase could catalyze the hydrolysis of L－amino acid esters, but not D－amino acid esters.
The C60－cryptand[2,2] coated piezoelectric (PZ) crystal sensor with immobilized C60－lipase was prepared to detect L－amino acid esters in aqueous solutions. The home－made optical isomer PZ sensor for esters was based on the adsorption of L－amino acid, the product of the hydrolysis of L－amino acid ester with C60－lipase, onto the C60－cryptand[2,2] coated quartz crystal. The optical isomer PZ sensor for esters based on C60－lipase was successfully to be applied to distinguish between D－form and L－form amino acid esters. The optical isomer PZ sensor for esters exhibited good sensitivity and reproducibility for L－amino acid esters in aqueous solutions.

並列關鍵字

fullerene ； lipase ； piezoelectric crystal sensor ； optical isomer