請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/3366
標題: Purification of Chitinase Using Peanibacillus sp. Culture Modified Chitin
應用Peanibacillussp.發酵修飾幾丁質進行幾丁酵素之純化研究
作者: Lin, Kai-Mine
林鎧民
關鍵字: 幾丁質
純化
幾丁酵素
出版社: 化學工程學系
摘要: The affinity chitin chromatography with fermentation-modified chitin was employed for purification of chitinase. For studying chitinase adsorption capacity, factors such as loading temperature, pH and elution solutions were investigated. The optimal loading conditions were found at 15℃, pH5, and elution conditions were: 0.02M acetic acid solution. Under this condition, the obtained specific chitinase activity gave a purification fold of 15.64. Furthermore, for investigating the mechanism of increase in the purification fold, it was found small particle size of chitin had a better adsorption capacity. Whereas, with the same chitin particle size of mesh 80, it was observed that enzyme-modified chitin gave an increase of 20.6% chitinase adsorption capacity and the fermentation-modified chitin yielded an increase of 54.2% chitinase adsorption capacity. By observing the structural changes of enzyme-modified and fermentation-modified chitin by SEM, it was found the chitin surface became porous & branching structure, which might reveal more active site & reduce the steric hinderance to the adsorption of chitinase.
本研究以經發酵後的幾丁質為載體利用幾丁質對幾丁酵素的親合性來進行純化,即為幾丁質親合層析。文中探討幾丁酵素吸附在幾丁質上之最適化條件,例如:酵素液載入時的溫度、pH,與幾丁質及膠態幾丁質來進行吸附能力的比較。從實驗得知在15℃、pH 5條件下可得到最高酵素吸附量,且吸附能力以經發酵修飾後的幾丁質較佳,使用沖提液以0.01M醋酸水溶液下為佳,所得之幾丁酵素純化倍數為15.64倍。 第二階段之研究內容為探討幾丁質的改質情形。實驗發現幾丁質的顆粒越小,其酵素吸附量越高;相同的幾丁質粒徑情況下,經酵素修飾後的幾丁質之酵素吸附量,較未修飾者高20.6%。而利用經發酵修飾後幾丁質酵素吸附量,則約可提升54.2%。以SEM圖觀察幾丁質經酵素及發酵修飾後的表面,發現幾丁質由平滑的表面變成多孔狀的結構,推測為幾丁質的直鏈結構,因此推知,經發酵修飾之幾丁質在吸附幾丁酵素時減少了立體障礙,進而提高對酵素的吸附量。
URI: http://hdl.handle.net/11455/3366
顯示於類別:化學工程學系所

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