Improved Stability of Soybean and Horseradish Peroxidases by Covalent Chemical Modification a
Improved Stability of Soybean and Horseradish Peroxidases by Covalent Chemical Modification a
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通过共价化学修饰提高大豆和辣根过氧化物酶的稳定性
DOI：
10.1111/j.1749-6632.1998.tb10371.x
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
P. Sundaram
中科院分区：
文献类型：
--
作者：
R. Venkatesh;P. Sundaram
方法辣根过氧化物酶和大豆过氧化物酶分别与分子量为100，1000和400 kDa的戊二醛（MGA），聚戊二醛（PGA）或氧化蔗糖聚合物（OSP）共价交联，在pH 8.0的50 mM磷酸钠缓冲液中进行还原脱氨程序。按照Tor等的方法，通过戊二醛聚合制备PGA。2用三硝基苯磺酸跟踪改性程度。用H2O2和ABTS法测定过氧化物酶活性. 3对天然和修饰的过氧化物酶进行了表征，并研究了温度、尿素和有机溶剂对它们的失活动力学的影响。在一段时间内测量热失活动力学，并使用失活速率常数（ki）计算半衰期和失活活化能。
METHODSHorseradish and soybean peroxidases were covalently cross-linked with glutaraldehyde (MGA), polyglutaraldehyde (PGA), or oxidized sucrose polymer (OSP) of molecular weights of 100, 1000, and 400 kDa, respectively, at pH 8.0 in 50 mM sodium phosphate buffer following the reductive deamination procedure. PGA was prepared by polymerizing glutaraldehyde according to the method of Tor et al. 2 The extent of modification was followed by using trinitrobenzene sulfonic acid. The activity of the peroxidase was assayed using H 2 O 2 and ABTS. 3 The native and modified peroxidases were characterized and their inactivation kinetics were studied under the influence of temperature, urea, and organic solvents. Thermal inactivation kinetics were measured over a period of time and the inactivation rate constant (k i) was used to calculate the half-life and the activation energy of inactivation.