Bioassessment of Acetylide-Imine in Acanthamoeba sp. Through In Vitro and In Silico Investigations for Its Environmental Contamination Implications from Electronic Waste

ASM Sc. J., 20(2), 2025

Published on December 9, 2025

https://doi.org/10.32802/asmscj.2025.0058

Author: Thivyan Manisekaran, Wan M. Khairul, Mas Mohamad, Yinn Dorng Foong, Eu Ling Hui, Aidatul Aifa Mohd Tajudin, Syed Ahmad Tajudin Tuan Johari, Siti Aisyah Razali, Fatimah Hashim, Rafizah Rahamathullah

Abstract

FYD4A, an organically synthesised molecule featuring an acetylide-imine motif in its molecular backbone, has potential as an electronic component. The compound’s safety was evaluated on the eukaryotic microorganism, Acanthamoeba sp., which is highly sensitive to environmental changes. Dose-response test using the MTT assay, and morphological observations via light and fluorescence microscopy, assess the cytoplasmic activity. Apoptosis confirmation was through DNA fragmentation analysis. Finally, in silico analysis examines the interaction between FYD4A and Acanthamoeba profilin II protein. The IC50 value indicates that FYD4A is moderately toxic to Acanthamoeba (9.15 µg/mL). FYD4A caused cell shrinkage and led to the encystment stage. Fluorescent staining revealed that the majority of the cells experienced membrane leakage, indicating signs of necrosis and random fragmentation in the DNA structure and smearing, which is another indicator of necrotic effects by FYD4A. Observation of FYD4A's interaction with Acanthamoeba profilin protein (II) showed strong binding to the following amino acids: Ile70, Arg71, Arg75, Tyr78, Pro106, Ala110, Glu114, Asp118, and Il e121. Understanding the detailed effects of synthetic compounds, particularly for electronic applications, is crucial, as improper disposal could lead to adverse environmental impacts, starting with their effects on microorganisms in the environment, such as Acanthamoeba.

Keywords: acetylide-imine, microscopy techniques, molecular docking, necrosis, protein