尿素酶
化学
固定化酶
共价键
Mercury(编程语言)
酶分析
尿素
自愈水凝胶
酶
丙烯酰胺
热稳定性
色谱法
核化学
聚合物
高分子化学
生物化学
有机化学
单体
程序设计语言
计算机科学
作者
Konda Reddy Kunduru,S. N. Raju Kutcherlapati,Dhamodaran Arunbabu,Tushar Jana
出处
期刊:Methods in Enzymology
日期:2017-01-01
卷期号:: 143-167
被引量:9
标识
DOI:10.1016/bs.mie.2017.02.008
摘要
Jack bean urease is an important enzyme not only because of its numerous uses in medical and other fields but also because of its historical significance—the first enzyme to be crystallized and also the first nickel metalloenzyme. This enzyme hydrolyzes urea into ammonia and carbon dioxide; however, the stability of this enzyme at ambient temperature is a bottleneck for its applicability. To improve urease stability, it was immobilized on different substrates, particularly on polymeric hydrogels. In this study, the enzyme was coupled covalently with poly(acrylamide) hydrogel with an yield of 18 μmol/cm3. The hydrogel served as the nanoarmor and protected the enzyme against denaturation. The enzyme immobilized on the polymer hydrogel showed no loss in activity for more than 30 days at ambient temperature, whereas free enzyme lost its activity within a couple of hours. The Michaelis–Menten constant (Km) for free and immobilized urease were 0.0256 and 0.2589 mM, respectively, on the first day of the study. The Km of the immobilized enzyme was approximately 10 times higher than that of the free enzyme. The hydrogel technique was also used to prepare light diffracting polymerized colloidal crystal array in which urease enzyme was covalently immobilized. This system was applied for the detection of mercury (Hg2 +) with the lower limit as 1 ppb, which is below the maximum contaminant limit (2 ppb) for mercury ions in water. The experimental details of these studies are presented in this chapter.
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