2005-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/704871摘要：幾丁質水解酵素ChiCW及ChiCH為拮抗細菌Bacillus cereus 28-9所產生，ChiCW具有明顯之抑菌活性；ChiCH僅具有微弱之抑菌活性。為獲得足夠量之酵素以進行抑菌及生化分析，擬利用大腸桿菌表現系統大量表現二幾丁質水解酵素，建立蛋白質表現系統及蛋白質純化之步驟，並進行最適作用溫度、酸鹼值、水解產物分析等，以對這兩種序列完全不同之幾丁質水解酵素在催化活性及酵素作用機制上有更深入的瞭解，初探二酵素抑菌能力差異之可能導因。<br> Abstract: Chitinase ChiCW and ChiCH are produced by antagonistic bacterium Bacillus cereus 28-9. ChiCW exhibits significant inhibitory activity against fungal spore germination, but ChiCH has only mild antifungal activity. To obtain enough amounts of proteins for antifungal and biochemical analysis, production of ChiCW and ChiCH will be tried by using Escherichia coli protein expression system. Protein purification procedure for these two chitinases will be established. Purified proteins will be used to analyze the optimal reaction temperature, pH, hydrolyzing products etc. to get better understanding on the catalytic activity and enzymatic mechanism of two chitinases, also the causes for the discrepancy of antifungal activity exhibited by these chitinases.幾丁質水解酵素ChiCWChiCH重組蛋白蛋白質純化chitinaseChiCWChiCHfusion protein protein purification