應用幾丁質及奈米複合顆粒於幾丁聚醣酶與蛋白酶之純化

Application of chitin particles on the purification of chitinolytic enzymes and proteases

陳政融(Jung-Long Chen)

指導教授 ： 王三郎

共同指導教授 ： 薛文發

淡江大學/理學院/化學學系碩士班/碩士(2011年)

https://doi.org/10.6846/TKU.2011.00692

摘要

使用幾丁類物質粗製蝦殼粉 (Crude SSC) 與磁性幾丁聚醣/四氧化三鐵納米複合顆粒 (NF) 以吸附方式純化酵素，以達成快速純化之目的。
由吸附劑SP、AC、pure chitin、Crude SSC、Crude CSC、Crude SPC中選擇較適吸附劑Crude SSC。再探討溫度、重量、時間對吸附鳳梨酵素之影響。經由實驗得知於4℃、30分鐘、0.1 g的條件下，得到比活性0.0918 U/mg。
再探討溫度、重量、時間對NF吸附鳳梨酵素之影響。從實驗得知於25℃、30分鐘、0.1 g 的條件，得到比活性0.1357 U/mg。
使用上述條件，使用NF與Crude SSC吸附細菌酵素，藉此討論吸附方式純化酵素的結果。經由實驗可知純化酵素影響條件並非只有等電點的因素，也需考慮酵素和吸附劑的相互作用還有物理吸附等因素。
結果證明吸附方式純化酵素是一個可行的純化方式，可以提高純化效率。

關鍵字

蛋白酶 ； 幾丁質酶 ； 磁性幾丁聚醣/四氧化三鐵納米複合顆粒 ； 粗製蝦殼幾丁質

並列摘要

Application of NF and Crude SSC on the purification of enzymes to achieve rapid purification purposes. Adsorbent : SP, AC, pure chitin, Crude SSC, Crude CSC, Crude SPC , Choose a more suitable adsorbent Crude SSC. Studying temperature, weight, time on the adsorption effect of the pineapple enzyme. By experiment that at 4 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.0918 U / mg.
Studying temperature, weight, time on the effect of NF adsorption bromelain enzyme. Learned from the experiment at 25 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.1357 U / mg.
Use the above conditions, NF, and Crude SSC absorption bacterial enzyme purified enzyme to discuss the results. Purified enzyme affected by the experimental conditions known isoelectric point is not the only factor, but also need to consider the interaction between enzyme and adsorbents as well as physical absorption and other factors.
Results show that absorption is a viable way to purify the enzymes, it can improve the purification efficiency.

並列關鍵字

Protease ； chitinase ； Magnetic chitosan / Fe3O4 nanocomposite particles ； Crude SSC

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