Evaluation of 2,6-dichlorophenolindophenol acetate as a substrate for acetylcholinesterase activity assay

Abstract

Ellman’s method is a standard protocol for the determination of cholinesterases activity. Though the method is ready for laboratory purposes, it has some drawbacks as well. In the current article, 2,6-dichloroindophenol acetate is performed as a chromogenic substrate suitable for acetylcholinesterase (AChE) activity examination. Michaelis constant and maximal velocity for 2,6-dichloroindophenol acetate were determined (38.0 µM and 244 pkat) and compared to the values for acetythiocholine (K_m 0.18 mM; V_max 5.1 nkat). Docking for 2,6-dichloroindophenol acetate and human AChE was done as well. In conclusion, 2,6-dichloroindophenol acetate seems to be suitable chromogenic substrate for AChE and spectrophotometry and based on this it can be easily performed whenever AChE activity should be tested.

• Acetylcholinesterase
• assay
• butyrylcholinesterase
• chromogen
• Ellman method
• spectrophotometry
• substrate

Acknowledgements

The European Union is gratefully acknowledged for project TEAB; CZ.1.07/2.3.00/20.0235. A long-term organization development plan 1011 (Faculty of Military Health Sciences, University of Defence, Czech Republic) is acknowledged as well.

Declaration of interest

The authors declare no conflicts of interests. The authors alone are responsible for the content and writing of this article.