biostudies-literatureLin XDrug safety research project of Guangxi Zhuang Autonomous Region Drug AdministrationGuangxi Natural Science Foundation2022 Guangxi Higher Education Institutions Young and Middle-aged Teachers Scientific Research Basic Ability Enhancement ProjectNational Natural Science Foundation of ChinaSelf-financed scientific research of Guangxi Zhuang Autonomous Region Health Commission in 2022Self-financed scientific research of Guangxi Zhuang Autonomous Region Health Commission1961https://www.ebi.ac.uk/biostudies/studies/S-EPMC11085145biostudies-literatureUnknown29(9)In this study, two "on-off" probes (BF₂-cur-Ben and BF₂-cur-But) recognizing acetylcholinesterase (AChE) were designed and synthesized. The obtained probes can achieve recognition of AChE with good selectivity and pH-independence with a linear range of 0.5~7 U/mL and 0.5~25 U/mL respectively. BF₂-cur-Ben has a lower limit of detection (LOD) (0.031 U/mL), higher enzyme affinity (K_m = 16 ± 1.6 μM), and higher inhibitor sensitivity. A responsive mechanism of the probes for AChE was proposed based on HPLC and mass spectra (MS) experiments, as well as calculations. In molecular simulation, BF₂-cur-Ben forms more hydrogen bonds (seven, while BF₂-cur-But has only four) and thus has a more stable enzyme affinity, which is mirrored bMolecules (Basel, Switzerland)Two Fluorescent Probes for Recognition of Acetylcholinesterase: Design, Synthesis, and Comparative Evaluation.PMC11085145GYJK (2023)0032022GXNSFAA035453Z-A20221011222670022022KY1387Qing BJiang JZou LJiang FHuang JLin XLiu QYi QLan WWang JWang MLai ZHuang XfalseTwo Fluorescent Probes for Recognition of Acetylcholinesterase: Design, Synthesis, and Comparative Evaluation.In this study, two "on-off" probes (BF₂-cur-Ben and BF₂-cur-But) recognizing acetylcholinesterase (AChE) were designed and synthesized. The obtained probes can achieve recognition of AChE with good selectivity and pH-independence with a linear range of 0.5~7 U/mL and 0.5~25 U/mL respectively. BF₂-cur-Ben has a lower limit of detection (LOD) (0.031 U/mL), higher enzyme affinity (K_m = 16 ± 1.6 μM), and higher inhibitor sensitivity. A responsive mechanism of the probes for AChE was proposed based on HPLC and mass spectra (MS) experiments, as well as calculations. In molecular simulation, BF₂-cur-Ben forms more hydrogen bonds (seven, while BF₂-cur-But has only four) and thus has a more stable enzyme affinity, which is mirrored b2024-01-01T00:00:00Z2024 Apr2026-04-24T03:16:00.677Z2026-04-24T03:10:05.581ZS-EPMC110851453873145210.3390/molecules29091961