Clostridium perfringens Sialidase Interaction with Neu5Ac α-Gal Sialic Acid Receptors by In- Silico Observation and Its Impact on Monolayers Cellular Behaviour Structure

Abstract

Bacterial sialidase hydrolyzes sialic acid receptors, preventing viral infections. Previous study reported Clostridium perfringens sialidase has inhibited viral infection in cell culture and animal models. Since the receptors have not been independently demonstrated, the effects of sialidase on host cells and their behavior are still poorly known. This study aims to evaluate C. perfringens sialidase based antiviral drugs using computational screening and in vitro approach to demonstrate interaction between enzyme-based drugs and ligand in host cells. Insilico study was carried out by molecular docking analysis used to predict the interactions between atoms that occur, followed by genetic characterization of sialidase from wild isolate. Sialidase which has undergone further production and purification processes exposed to chicken embryonic fibroblast cell culture, and observations based structural morphology of cell compared between treated cell and normal cell without treatment. Based on in-silico study shows that C. perfringens sialidase have an excellent binding affinity with Neu5Acα (2.3) Gal ligand receptor with Gibbs energy value (∆G) − 7.35 kcal/mol and Ki value of 4.11 µM. Wild C. perfringens isolate in this study have 99.1–100% similarity of plc gene, NanH, and NanI genes, while NanJ shows 93.18% similarity compared to reference isolate from GenBank. Sialidase at 750 mU and 150 mU may impact on the viability, cell count, and cell behavior structure of fibroblast cells by significance increased the empty area and perimeter of CEF cells, while at 30 mU sialidase shows no significant difference compared with mock control. Sialidase derived C. perfringens has the capacity to compete with viral molecules for attachment to hosts sialic acid based on in-silico analysis. However, sialidase treatment cause an impact on monolayer cell fibroblasts given exposure to high doses.