Mahmoudi Niloofar, Fatemi Fataneh, Rahmandoust Moones, Mirzajani Fateme, Ranaei Siadat Seyed Omid
Protein Research Center, Shahid Beheshti University, Tehran, Iran.
Heliyon. 2023 Aug 30;9(9):e19551. doi: 10.1016/j.heliyon.2023.e19551. eCollection 2023 Sep.
In this study, a proper and reliable fluorometric method is introduced for screening acetylcholinesterase (AChE) and its inhibitors, using carbon quantum dots (CQDs) as the signal reporter. Pure, S-doped, and P-doped CQDs, were synthesized and their recoverable fluorescence quenching properties were observed, when exposed to Hg, Cu, and Fe quenching ions, respectively. The study on the recovery of their emission showed that after the introduction of another guest substance with a stronger affinity to the quenching ions, their fluorescence is restored. The Design Expert software was employed to compare the performance of the three CQDs, as fluorescent probes, based on their quenching efficiency and the percentage of their emission recovery in the presence of AChE and acetylthiocholine (ATCh). Based on the statistical analysis, among the studied CQDs, S-doped CQD was the most suitable candidate for sensor designing. The detection mechanism for the proposed S-doped CQD-based sensor is as follows: The strong binding of Cu ions to carboxyl groups of S-doped CQD quenches the fluorescence signal. Then, hydrolysis of ATCh into thiocholine (TCh) in the presence of AChE causes fluorescence recovery, due to the stronger affinity of Cu to the TCh, rather than the CQD. Finally, in the presence of malathion and chlorpyrifos inhibitors, AChE loses its ability to hydrolyze ATCh to TCh, so the fluorescence emission remains quenched. Based on the proposed detection technique, the designed sensor showed detection limits of 1.70 ppb and 1.50 ppb for malathion and chlorpyrifos, respectively.
在本研究中,介绍了一种合适且可靠的荧光法,用于筛选乙酰胆碱酯酶(AChE)及其抑制剂,该方法使用碳量子点(CQDs)作为信号报告分子。合成了纯的、硫掺杂的和磷掺杂的碳量子点,并分别观察到它们在暴露于汞、铜和铁猝灭离子时具有可恢复的荧光猝灭特性。对其发射恢复的研究表明,在引入另一种对猝灭离子具有更强亲和力的客体物质后,它们的荧光得以恢复。使用Design Expert软件,基于三种碳量子点作为荧光探针的猝灭效率以及在乙酰胆碱酯酶(AChE)和乙酰硫代胆碱(ATCh)存在下其发射恢复的百分比,对它们的性能进行了比较。基于统计分析,在所研究的碳量子点中,硫掺杂的碳量子点是传感器设计的最合适候选物。所提出的基于硫掺杂碳量子点的传感器的检测机制如下:铜离子与硫掺杂碳量子点的羧基强烈结合,使荧光信号猝灭。然后,在乙酰胆碱酯酶存在下,乙酰硫代胆碱水解为硫代胆碱(TCh),由于铜对硫代胆碱的亲和力强于对碳量子点的亲和力,导致荧光恢复。最后,在马拉硫磷和毒死蜱抑制剂存在下,乙酰胆碱酯酶失去将乙酰硫代胆碱水解为硫代胆碱的能力,因此荧光发射保持猝灭。基于所提出的检测技术,所设计的传感器对马拉硫磷和毒死蜱的检测限分别为1.70 ppb和1.50 ppb。