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yungbonjut

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1.5 脂肪酶固定化
采用单因素对比试验,对使用不同浓度AgNO3制备出的Ag-MAC进行比较选择,对固载的pH环境和固载时间进行优化实验。
取2mg/mL的酶溶液20mL、Ag-MAC40mg,加入50mL圆底离心管中,混合液在25℃,215rpm的条件下,进行酶固定化。在混合液反应到一定时间后,取出离心管在10℃,6000rpm条件下,离心10min。用考马斯亮蓝蛋白方法测定上清液中蛋白量,上清液中减少的蛋白量即是Ag-MAC上所固定的酶蛋白量,固定化的酶蛋白与载体量的比例可以由下面的计算式得出:

其中,Pa是单位载体上固定的酶蛋白的量(mg/g),Ci和Cf反应前后酶溶液的浓度(mg/mL),V是反应液体积(mL);W是使用的载体的重量(mg)。

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爱与雨下(金币+1): 2011-11-04 14:17:47
爱与雨下(金币+40): bb代发 2011-11-10 08:50:15
1.5 Immobilization of lipase.
Via single factor comparison test, compare and exam Ag-MAC produced by different concentrations of AgNO3 and optimize test on pH environment and time of immobilization.
Select 20mL of enzyme solution (2mg/mL) and 40mg of Ag-MAC and add them to 50mL round centrifuge tube. Immobilize enzyme is processed by mixed solution under condition of 25C and 215rpm. After mixture reacting for certain time, take it out and centrifuge for 10min under 10C and 6000rpm. Test protein contents in nascent by coomassie blue staining. The reduced amount of protein in nascent is amount immobilized protein and the ratio of immobilized protein and carrier can be obtained by following formula.
In formula, Pa is amount of immobilized enzyme protein on unit carrier (mg/g), Ci and Cf are concentrations of solution before and after reaction (mg/mL), V is reacted solution volume (mL), W is mass of used carrier (mg).
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