Regulation of Proteolytic Activity to Improve the Recovery of Macrobrachium rosenbergii Nodavirus Capsid Protein

The causative agent of white tail disease (WTD) in the giant freshwater prawn is Macrobrachium rosenbergii nodavirus (MrNV). recombinant capsid protein (CP) MrNV was previously expressed Escherichia coli, and it self-assembled into icosahedral virus-like particles (VLPs) with a diameter approximately 30 nm. Extensive studies on CP VLPs have attracted widespread attention their potential applications as biological nano-containers for targeted drug delivery antigen display scaffolds vaccine developments. Despite advantageous features, produced E. coli are seriously affected by protease degradations, which significantly affect yield stability VLPs. Therefore, aim this study to enhance modulating degradation activity. Edman amino acid sequencing revealed that proteolytic cleavage occurred at arginine 26 CP. proteases responsible were predicted silico using Peptidecutter, Expasy. To circumvent proteolysis, specific inhibitors (PMSF, AEBSF E-64) tested reduce rates. Modulation activity demonstrated cysteine degradation. addition E-64, inhibitor, remarkably improved 2.3-fold compared control. This innovative approach generates an economical method improve scalability individual inhibitors, enabling retain structural integrity prominent downstream including development.