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肌紅蛋白熱變性之旋光量測研究
Thesis

肌紅蛋白熱變性之旋光量測研究

林志龍
Masters, 國立清華大學, 生醫工程與環境科學系
2014

Abstract

肌紅蛋白 旋光 Myoglobin Optical Rotation
Recently, many measurement methods have been used to study structural changes of proteins after heating, such as differential scanning calorimetry method, circularly polarized dichroism spectroscopy, X-ray diffraction crystallography method, etc. However, it is difficult for these methods to monitor the denaturation process of myoglobin solution in real time. In this study, a self-assembled optical heterodyne polarimeter capable of amplifying the optical rotation signal to 41-fold, and a precision thermoelectric cooler (TEC) were used to study the thermal denaturation phenomenon of myoglobin solution after heating. Our results indicate that the protein structure of myoglobin solution has about 5% destroyed by heating to 75℃, however, by cooling down, the protein structure is reversible at this time because of protein renaturation role. When myoglobin solution was heated to about 75.9±0.1°C,its protein structure was gradually changed from reversible to partially reversible. As the myoglobin solution was heated to 77.5℃ and then cooled down to room temperature, about 80% of proteins reversed to original protein structure. Furthermore, as the myoglobin was heated to 80℃ and 85℃, there was 52% and 45%, respectively, proteins to be reversible to the original form. The helical structure of myoglobin were mostly destroyed between 75.9 and 80.4℃. Upon cooling to 25℃ from tempertures above 80.4℃,the recovered helical structure decreased with rise of temperature before cooling.

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