Topcular C, Ayhan H
Hacettepe University, Institute of Pure and Applied Science, Bioengineering Division, Beytepe, Ankara, Turkey.
J Biomater Sci Polym Ed. 2007;18(5):595-607. doi: 10.1163/156856207780852550.
The aim of this study was to immobilize horseradish peroxidase (HRP) onto poly(glycidyl methacrylate) (PGMA) microspheres. In this study, PGMA particles were synthesised by dispersion polymerization from the glycidyl methacrylate (GMA) monomer. The monomer conversion value was determined to be 89% by weight. The first step comprised opening of the epoxide rings and formation of amine groups on the particle surfaces. The characterised particles were used as micro-carriers for the immobilisation of HRP. Glutaraldehyde, as bifunctional reactive, has been used as the spacer arm in the second step of the covalent binding method. The optimum immobilisation conditions found are as follows: immobilisation time 10 min, pH 7.2, temperature 25 degrees C, initial enzyme concentration 0.05 mg/ml. Kinetic parameters of native and immobilised enzyme, such as K(m) and V(max) values, have been calculated 2.33 mmol/l and 0.670 mmol/l per s for the native enzyme and for the immobilised enzyme 69.13 mmol/l and 0.175 mmol/l per s, respectively. The HRP immobilised microparticules were used to purify a textile bleaching solution in which excess H(2)O(2) remains.
本研究的目的是将辣根过氧化物酶（HRP）固定在聚甲基丙烯酸缩水甘油酯（PGMA）微球上。在本研究中，PGMA颗粒由甲基丙烯酸缩水甘油酯（GMA）单体通过分散聚合反应合成。单体转化率经重量法测定为89%。第一步包括环氧环的开环以及在颗粒表面形成胺基。经表征的颗粒用作固定HRP的微载体。戊二醛作为双功能反应剂，在共价结合法的第二步中用作间隔臂。所发现的最佳固定条件如下：固定时间10分钟、pH值7.2、温度25℃、初始酶浓度0.05mg/ml。已计算出天然酶和固定化酶的动力学参数，如K(m)和V(max)值，天然酶分别为2.33mmol/l和0.670mmol/(l·s)，固定化酶分别为69.13mmol/l和0.175mmol/(l·s)。固定有HRP的微粒用于纯化残留过量H(2)O(2)的纺织品漂白溶液。
