Khalife Majd, Stankovic Dalibor, Stankovic Vesna, Danicka Julia, Rizzotto Francesco, Costache Vlad, Schwok Anny Slama, Gaudu Philippe, Vidic Jasmina
Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, 78350 Jouy en Josas, France.
Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia.
Food Chem. 2024 Feb 1;433:137316. doi: 10.1016/j.foodchem.2023.137316. Epub 2023 Aug 31.
Vitamin K refers to a group of vitamins that play an important role in blood coagulation and regulation of bone and vascular metabolism. However, vitamin K may give severe side effects in animal and humans when improperly added to food and feed due to its toxicity. Here, an electrochemical biosensor, based on the YaiB NADPH-dependent quinone reductase from Lactococcus lactis (YaiB), was developed to achieve rapid and redox probe-free detection of vitamin K. First, the ability of the carbon electrode to distinguish between 1,4-benzoquinone and hydroquinone was demonstrated. Then, we engineered YaiB to work as a bioreceptor immobilized at the electrode and its sensitivity and specificity to reduce vitamin K were demonstrated. Finally, to demonstrate the practical potential of the biosensor, we tested it directly in spiked milk samples, achieving 15-minute quantification of the vitamin K. The limit of detection was 0.87 µM and 4.1 µM in buffer and milk, respectively.
维生素K是指一组在血液凝固以及骨骼与血管新陈代谢调节中发挥重要作用的维生素。然而，由于其毒性，维生素K在不当添加到食品和饲料中时，可能会对动物和人类产生严重的副作用。在此，基于乳酸乳球菌的YaiB烟酰胺腺嘌呤二核苷酸磷酸（NADPH）依赖性醌还原酶（YaiB）开发了一种电化学生物传感器，以实现对维生素K的快速且无需氧化还原探针的检测。首先，证明了碳电极区分1,4-苯醌和对苯二酚的能力。然后，我们对YaiB进行改造，使其作为固定在电极上的生物受体，并证明了其还原维生素K的灵敏度和特异性。最后，为了证明该生物传感器的实际应用潜力，我们直接在加标的牛奶样品中对其进行测试，实现了对维生素K的15分钟定量测定。在缓冲液和牛奶中的检测限分别为0.87 μM和4.1 μM。
