Department of Comparative Biomedicine and Food Science, University of Padua, Agripolis-Viale dell'Università 16, 35020, Legnaro, PD, Italy.
Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacký University in Olomouc, 17 Listopadu 12, 771 46, Olomouc, Czech Republic.
Amino Acids. 2019 Apr;51(4):679-690. doi: 10.1007/s00726-019-02704-7. Epub 2019 Feb 6.
The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein-nanoparticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanoparticles (SAMNs) represent a new class of naked magnetic nanoparticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV-Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.
保持酶的活性对于开发杂交纳米-生物缀合物至关重要,而控制蛋白质-纳米颗粒相互作用,从而形成稳定且具有催化活性的杂化体,则是设计新型生物传感平台的关键。在这种情况下,表面活性磁赤铁矿纳米粒子(SAMNs)代表了一类新型的裸露磁性纳米粒子,具有独特的电催化特性和选择性结合蛋白质的能力。番茄重组氨基酸醛脱氢酶(SlAMADH1)被用作模型蛋白,并通过自组装成功固定在裸露的 SAMNs 表面,其酶活性在超过 6 个月的时间里得以保持。通过紫外可见光谱、质谱和 TEM 显微镜对混合纳米材料(SAMN@SlAMADH1)进行了表征,并将其应用于开发用于测定酒精饮料中氨基酸醛的生物传感器。在包含修饰有 SAMN 的碳糊电极的低体积电化学流动池中进行测量,用于库仑法测定酶催化过程中产生的 NADH。本研究不仅代表了在酒精基质中用于氨基酸醛的电化学生物传感器的首例,而且为在裸露金属氧化物纳米材料上固定酶用于生物传感开辟了道路。