Šekuljica Nataša Ž, Prlainović Nevena Ž, Jovanović Jelena R, Stefanović Andrea B, Djokić Veljko R, Mijin Dušan Ž, Knežević-Jugović Zorica D
Faculty of Technology and Metallurgy, Innovation Center, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Republic of Serbia.
Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000, Belgrade, Republic of Serbia.
Bioprocess Biosyst Eng. 2016 Mar;39(3):461-72. doi: 10.1007/s00449-015-1529-x. Epub 2016 Jan 8.
Kaolin showed as a very perspective carrier for the enzyme immobilization and it was used for the adsorption of horseradish peroxidase (HRP). The effects of the enzyme concentration and pH on the immobilization efficiency were studied in the reaction with pyrogallol and anthraquinone dye C.I. Acid Violet 109 (AV 109). In addition, Fourier transform infrared spectroscopy, scanning electron microscopy and analysis by Brunauer-Emmett-Teller were performed for kaolin, thermally activated kaolin and the immobilized enzyme. It has been shown that 0.1 IU of HRP-kaolin decolorized 87 % of dye solution, under the optimal conditions (pH 5.0, temperature 24 °C, dye concentration 40 mg/L and 0.2 mM of H2O2) within 40 min. The immobilized HRP decolorization follows the Ping Pong Bi-Bi mechanism with dead-end inhibition by the dye. The biocatalyst retained 35 ± 0.9 % of the initial activity after seven cycles of reuse in the decolorization reaction of AV 109 under optimal conditions in a batch reactor. The obtained kinetic parameters and reusability study confirmed improvement in performances of k-HRP compared to free, indicating that k-HRP has a great potential for environmental purposes.
高岭土是一种极具前景的酶固定化载体，被用于吸附辣根过氧化物酶（HRP）。在与邻苯三酚和蒽醌染料C.I.酸性紫109（AV 109）的反应中，研究了酶浓度和pH对固定化效率的影响。此外，对高岭土、热活化高岭土和固定化酶进行了傅里叶变换红外光谱、扫描电子显微镜和Brunauer-Emmett-Teller分析。结果表明，在最佳条件（pH 5.0、温度24℃、染料浓度40 mg/L和0.2 mM H2O2）下，0.1 IU的HRP-高岭土在40分钟内可使87%的染料溶液脱色。固定化HRP的脱色遵循乒乓双底物机制，染料会产生终产物抑制。在间歇反应器中，在最佳条件下对AV 109进行脱色反应，经过七次重复使用后，该生物催化剂保留了初始活性的35±0.9%。所获得的动力学参数和可重复使用性研究证实，与游离酶相比，固定化HRP的性能有所提高，表明固定化HRP在环境应用方面具有巨大潜力。
