CAJ | 학술논문

利用溶胶-凝胶法将辣根过氧化物酶(HRP)固定化于二氧化硅凝胶网络中构建了可用于酚类化合物检测的酶传感器.对二氧化硅载体材料进行了结构表怔.二氧化硅多孔材料的平均孔径为2.95 nm.孔径小于5 nm占总数的84.068%.由于辣根过氧化物酶的分子尺寸远远大于二氧化硅的凝胶网络的平均孔径,因此不会泄露到溶液中去,而尺寸较小的底物可以发生反应.包埋的辣根过氧化物酶在H_2O_2的存在下,能够催化氧化苯酚与4-氨基安替比林反应生成醌亚胺有色化合物.通过紫外-可见光谱测定醌哑胺有色化合物的吸光度,即可以确立苯酚的含量.对于象含氯酚类的重要污染物,如邻氯酚、问氯酚、2,4-二氯酚,这种方法也同样适用.此外,对多次测定以后的酶的活性下降的问题进行了讨论,结果表明酶传感器可以重复使用10次以上.但响应时间会变长.
이용용효-응효법장랄근과양화물매(HRP)고정화우이양화규응효망락중구건료가용우분류화합물검측적매전감기.대이양화규재체재료진행료결구표정.이양화규다공재료적평균공경위2.95 nm.공경소우5 nm점총수적84.068%.유우랄근과양화물매적분자척촌원원대우이양화규적응효망락적평균공경,인차불회설로도용액중거,이척촌교소적저물가이발생반응.포매적랄근과양화물매재H_2O_2적존재하,능구최화양화분분여4-안기안체비림반응생성곤아알유색화합물.통과자외-가견광보측정곤아알유색화합물적흡광도,즉가이학립분분적함량.대우상함록분류적중요오염물,여린록분、문록분、2,4-이록분,저충방법야동양괄용.차외,대다차측정이후적매적활성하강적문제진행료토론,결과표명매전감기가이중복사용10차이상.단향응시간회변장.
Phenolic analysis was established on the immobilized horseradish peroxidase(HRP)catalyzed oxidation reaction.It subsequently catalyzed oxidative coupling of phenol with 4-aminophenazone using aqueous hydrogen peroxide to form intensely colored products for spectrophotometric analysis.HRPs were trapped in sol-gel matrix in a mild procedure.The immobilized HRPs maintain almost identical enzymatic activity as those in solution.BET analysis indicates that the silica itself is a porous structure with the average pore diameter of 2.95 nm.It permits small size molecules i.e.hydrogen peroxide,phenol and 4-aminophenazone to diffuse into the matrix while large molecules like enzyme(HRP)remain in the pores,It thus allows a biocatalysis to occur and makes the most out of the enzymes encapsulated in the silica matrix to stand against leakage so that the immobilized horseradish peroxidase could be recycled.The method can be employed for 2-ehlorophenol,3-chloropbenol or 2,4-dichlorophenol analysis as well.The feasibility of recycle on immobilized enzyme is evaluated.Although enzymatic activities are obviously decreased after repeated utilization of 9 times,the method definitely offers a potential spectrophotometric biosensor for phenolic compounds analysis.