固定化酶法测定微量组分  

Determination of Trace Component by Immobilized Enzyme

作　　者：宁静[1] 柳畅先[1] 

机构地区：[1]中南民族大学化学与材料科学学院,分析化学国家民委重点实验室,湖北武汉430074

出　　处：《分析科学学报》2015年第2期261-264,共4页Journal of Analytical Science

基　　金：中南民族大学自然科学基金(XTZ09005)

摘　　要：以壳聚糖为载体对醇脱氢酶(ADH)和脲酶(UASE)进行固定化。固定化的最适条件:ADH和UASE的偶联时间分别为2.5h和1.5h,交联剂戊二醛浓度为0.6%和0.5%,偶联pH值为6.8和5.0。对游离酶及固定化酶的性质研究表明,ADH酶促反应的最适宜pH为8.0和8.4,最适宜温度为33℃和35℃,米氏常数为48mmol/L和13mmol/L;UASE酶促反应的最适宜pH均为7.0,最适宜温度为60℃和72℃,米氏常数为0.4mmol/L和2mmol/L。固定化酶与游离酶相比在复用性上具有优势,用固定化酶测定了试样中尿素含量以及模拟样中银离子含量。Alcohol dehydrogenase（ADH）and urease（UASE）were immobilized on chitosan by covalant conjugation.The optimum conditions of immobilization for ADH and UASE were that chitosan was crosslinked with 0.6% and 0.5% glutaraldehyde,and reacted with enzymes for 2.5hand 1.5hin the solution of pH =6.8and 5.0,respectively.The enzymatic characterizations of free and immobilized enzymes were studied.For ADH,the optimum pH values of the enzymatic reaction were 8.0and 8.4,the temperatures were 33 ℃ and 35 ℃,the Kmvalues were 48 mmol/L and 13 mmol/L,respectively.For UASE,the optimum pH values of the enzymatic reaction were 7.0both,the temperatures were 60℃and72℃,the Kmvalues were 0.4mmol/L and 2mmol/L,respectively.It indicated that the immobilized enzyme had advantage of reuse stability over free enzyme.The immobilized enzyme has been used to determine the content of urea and silver ion in samples.

关 键 词：醇脱氢酶 脲酶 固定化 酶法测定 

分 类 号：O657.3[理学—分析化学]