碳六十脂肪酵素/三酸甘油酯壓電晶體感測器研製與應用

Preparation and Application of Piezoelectric Crystal Sensor for Triglyceride Based on C60-Lipase Enzyme

錢培宜(Pei-Yi Chien)

指導教授 ： 施正雄

國立臺灣師範大學/理學院/化學系/碩士(2006年)

摘要

碳六十脂肪酵素/三酸甘油酯壓電晶體感測器研製與應用

摘要

本研究合成碳六十-脂肪分解酵素(C60-lipase)，並應用在三酸甘油酯壓電晶體生物感測器。首先，利用塗佈碳六十的石英壓電晶體感測器作為研究碳六十與脂肪分解酵素間作用力，結果顯示酵素與碳六十存在不可逆的化學性吸附現象。且研究顯示碳六十與脂肪分解酵素間作用的最適溫度為32 ℃，最適pH值為7。
IR光譜圖利用來鑑定合成的不溶性碳六十-脂肪分解酵素(C60-lipase)。然後，使用多孔性的二氧化矽材質濾片固定化碳六十-脂肪分解酵素，這碳六十-脂肪分解酵素濾片顯示可用來催化水解三酸甘油酯溶液，造成pH值的下降。
本研究組裝一塗佈碳六十/大環胺醚的石英壓電晶體感測器用來吸附三酸甘油酯溶液經碳六十-脂肪分解酵素催化後的產物-脂肪酸，吸附所引起的負載重量增加會造成石英晶體振盪頻率的下降，並由電腦螢幕紀錄整個催化反應的過程。本研究對三乙酸甘油酯、三丁酸甘油酯及混合三酸甘油酯溶液進行偵測，分別探討了濃度、pH值效應及人體常見生物干擾物存在下的催化反應。結果顯示，此系統對三酸甘油酯的偵測具有良好的濃度線性關係及偵測極限。三乙酸甘油酯、三丁酸甘油酯及混合三酸甘油酯溶液其偵測極限依次為13.08 mg/dL、38.32 mg/dL、6.21 mg/dL皆低於人體內三酸甘油酯正常平均值50~150 mg/dL。同時，此利用C60-lipase而塗佈C60-cryptand[2.2]之生化感測器對於三乙酸甘油酯的偵測具有良好的再現性，RSD值為1.7 % (n= 5)。

關鍵字

三酸甘油酯 ； 脂肪酵素 ； 碳六十 ； 壓電晶體

並列摘要

Preparation and Application of Piezoelectric Crystal Sensor for Triglyceride Based on C60-Lipase Enzyme

Abstract

Immobilized fullerence C60-lipase was synthesized and applied in a piezoelectric crystal biosensor for triglycerides. Firstly, The C60 coated piezoelectric quartz crystal sensor was employed to study the interaction between C60 and lipase. It seemed to indicate that the lipase could be adsorbed on C60 by chemisorption. The optimum temperature of 32 ℃ and the optimum pH of 7 for the chemisorption between the C60 and lipase were found.
The water insoluble C60-lipase product was synthesized and identified by IR spectrum. The C60-lipase coated SiO2 filter was also prepared. The C60-lipase filter exhibited the catalytic ability for the hydrolysis of the triglycerides resulting in the decrease in pH of solutions.
The C60/cryptand[2.2] coated piezoelectric crystal biosensor was prepared for fatty acids , the product of the hydrolysis of the triglycerides catalyzed by C60-lipase. The adsorption of fatty acids on quartz crystal resulted in decrease in the oscillating frequency of quartz crystal. And the overall reaction process can be monitored and recorded by computer. The triglycerides e.g. triacetin, tributyrin and the mixed triglycerides were detected by the piezoelectric biosensor in this study. The effects of concentration, pH and the interference of the common biological species in human blood were also investigated. The piezoelectric sensor has exhibited good concentration linear relation and good detection limit for triglycerides. The detecting limits of the triacetin, tribuytrin and the mixed triglycerides were 13.08, 38.32 and 6.21 mg/dL, respectively which are all below the normal average value of the triglycerides in human body. The C60-cryptand[2.2] coated piezoelectric biosensor with C60-lipase also showed good reproducibility for reusage with RSD = 1.7 % (n = 5) for triglycerides.

參考文獻

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