Department of Analytical Biotechnology, Institute of Cell Biology National Academy of Sciences of Ukraine (ICB NASU), 14/16, Dragomanova Str., 79005 Lviv, Ukraine.
NanoTechnas-Center of Nanotechnology and Materials Science, Institute of Chemistry, Faculty of Chemistry and Geosciences, Vilnius University, Naugarduko 24, LT-03225 Vilnius, Lithuania.
Biosensors (Basel). 2023 May 28;13(6):587. doi: 10.3390/bios13060587.
L-Lactate is an indicator of food quality, so its monitoring is essential. Enzymes of L-Lactate metabolism are promising tools for this aim. We describe here some highly sensitive biosensors for L-Lactate determination which were developed using flavocytochrome b (Fc) as a bio-recognition element, and electroactive nanoparticles (NPs) for enzyme immobilization. The enzyme was isolated from cells of the thermotolerant yeast The possibility of direct electron transfer from the reduced form of Fc to graphite electrodes has been confirmed, and the amplification of the electrochemical communication between the immobilized Fc and the electrode surface was demonstrated to be achieved using redox nanomediators, both bound and freely diffusing. The fabricated biosensors exhibited high sensitivity (up to 1436 A·M·m), fast responses, and low limits of detection. One of the most effective biosensors, which contained co-immobilized Fc and the hexacyanoferrate of gold, having a sensitivity of 253 A·M·m without freely diffusing redox mediators, was used for L-Lactate analysis in samples of yogurts. A high correlation was observed between the values of analyte content determined using the biosensor and referenced enzymatic-chemical photometric methods. The developed biosensors based on Fc-mediated electroactive nanoparticles can be promising for applications in laboratories of food control.
L-乳酸是食品质量的指标,因此对其进行监测至关重要。L-乳酸代谢的酶是实现这一目标的有前途的工具。我们在这里描述了一些使用黄素细胞色素 b (Fc) 作为生物识别元件,以及用于酶固定化的电活性纳米颗粒 (NPs) 开发的高灵敏度 L-乳酸测定生物传感器。该酶是从耐热酵母细胞中分离出来的,已经证实还原型 Fc 可以与石墨电极直接进行电子转移,并且使用结合和自由扩散的氧化还原纳米介体证明可以实现固定化 Fc 与电极表面之间的电化学通信的放大。所制备的生物传感器具有高灵敏度(高达 1436 A·M·m)、快速响应和低检测限。其中一个最有效的生物传感器,含有共固定化的 Fc 和金的六氰合铁酸盐,在没有自由扩散的氧化还原介体的情况下具有 253 A·M·m 的灵敏度,用于酸奶样品中的 L-乳酸分析。使用生物传感器和参考酶化学比色法确定的分析物含量之间观察到高度相关性。基于 Fc 介导的电活性纳米颗粒的开发的生物传感器在食品控制实验室中具有广阔的应用前景。