Glycine as a stabilizing osmolyte for Renilla luciferase: A kinetic and molecular dynamics analysis

Abstract

Renilla luciferase, a luminescent enzyme, is utilized in gene expression analysis and biosensor technology, and extensive research has been conducted on its structure and function. Glycine is an osmolyte that plays a key role in protein stabilization against denaturation. However, its impact on enzyme properties is unpredictable. This study aimed to investigate the effect of glycine on the kinetics and stability of Renilla luciferase. The data revealed that glycine at a concentration range of 0.1 to 0.6 M improved enzyme kinetics and thermal stability. The highest catalytic efficiency was observed at a concentration of 0.5 M. Molecular dynamics simulations demonstrated that in the presence of 0.5 M glycine, substrate access to the enzyme’s active site was enhanced while the root-mean-square fluctuation (RMSF) of the protein backbone was reduced. Additionally, the analysis of protein-water hydrogen bonding interactions showed an increase in the hydrogen bonding between water molecules and Renilla luciferase. The present study may be used for the formulation of Renilla luciferase for commercial purposes. © 2024 University of Isfahan. Published by Informa UK Limited, trading as Taylor & Francis Group.