Department of Chemistry, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Pathumthani 12110, Thailand.
Division of Physics, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Pathumthani 12110, Thailand.
Biosensors (Basel). 2017 Oct 15;7(4):47. doi: 10.3390/bios7040047.
A biosensor based on chitosan-gelatin composite biopolymers nanofibers is found to be effective for the immobilization of horseradish peroxidase to detect hydrogen peroxide. The biopolymer nanofibers were fabricated by an electrospining technique. Upon optimization of synthesis parameters, biopolymers nanofibers, an average of 80 nm in diameter, were obtained and were then modified on the working electrode surface. The effects of the concentration of enzyme, pH, and concentration of the buffer and the working potential on the current response of the nanofibers-modified electrode toward hydrogen peroxide were optimized to obtain the maximal current response. The results found that horseradish peroxidase immobilization on chitosan-gelatin composite biopolymer nanofibers had advantages of fast response, excellent reproducibility, high stability, and showed a linear response to hydrogen peroxide in the concentration range from 0.1 to 1.7 mM with a detection limit of 0.05 mM and exhibited high sensitivity of 44 µA∙mMcm. The developed system was evaluated for analysis of disinfectant samples and showed good agreement between the results obtained by the titration method without significant differences at the 0.05 significance level. The proposed strategy based on chitosan-gelatin composite biopolymer nanofibers for the immobilization of enzymes can be extended for the development of other enzyme-based biosensors.
基于壳聚糖-明胶复合生物聚合物纳米纤维的生物传感器被发现可有效用于辣根过氧化物酶的固定化以检测过氧化氢。该生物聚合物纳米纤维是通过静电纺丝技术制备的。通过优化合成参数,获得了平均直径为 80nm 的生物聚合物纳米纤维,然后将其修饰在工作电极表面上。优化了酶浓度、pH 值、缓冲液浓度和工作电位对纳米纤维修饰电极对过氧化氢电流响应的影响,以获得最大的电流响应。结果发现,辣根过氧化物酶固定在壳聚糖-明胶复合生物聚合物纳米纤维上具有快速响应、优异的重现性、高稳定性的优点,并且在 0.1 至 1.7mM 的浓度范围内对过氧化氢表现出线性响应,检测限为 0.05mM,具有 44µA∙mMcm 的高灵敏度。所开发的系统用于消毒剂样品的分析,通过滴定法获得的结果之间具有良好的一致性,在 0.05 的显著性水平下没有显著差异。基于壳聚糖-明胶复合生物聚合物纳米纤维的酶固定化的这种策略可扩展到其他基于酶的生物传感器的开发。