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). The recombinant capsid protein (CP) of MrNV was previously expressed in Escherichia coli, and it self-assembled into icosahedral virus-like particles (VLPs) with a diameter of...
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MDPI
2021
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| Online Access: | http://eprints.sunway.edu.my/1835/ http://eprints.sunway.edu.my/1835/1/Yong%20Chean%20Yeah%20Regulation%20of%20proteolytic%20activity%20to%20improve%20the-ijms-22-08725.pdf |
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| author | Bethilda, A. S. Abdul Razak, M. Ho, K. L. Ng, C. L. Yong, Chean Yeah * Tan, W. S. |
| author_facet | Bethilda, A. S. Abdul Razak, M. Ho, K. L. Ng, C. L. Yong, Chean Yeah * Tan, W. S. |
| author_sort | Bethilda, A. S. |
| building | SU Institutional Repository |
| collection | Online Access |
| description | The causative agent of white tail disease (WTD) in the giant freshwater prawn is Macrobrachium rosenbergii nodavirus (MrNV). The recombinant capsid protein (CP) of MrNV was previously expressed in Escherichia coli, and it self-assembled into icosahedral virus-like particles (VLPs) with a diameter of approximately 30 nm. Extensive studies on the MrNV CP VLPs have attracted widespread attention in their potential applications as biological nano-containers for targeted drug delivery and antigen display scaffolds for vaccine developments. Despite their advantageous features, the recombinant MrNV CP VLPs produced in E. coli are seriously affected by protease degradations, which significantly affect the yield and stability of the VLPs. Therefore, the aim of this study is to enhance the stability of MrNV CP by modulating the protease degradation activity. Edman degradation amino acid sequencing revealed that the proteolytic cleavage occurred at arginine 26 of the MrNV CP. The potential proteases responsible for the degradation were predicted in silico using the Peptidecutter, Expasy. To circumvent proteolysis, specific protease inhibitors (PMSF, AEBSF and E-64) were tested to reduce the degradation rates. Modulation of proteolytic activity demonstrated that a cysteine protease was responsible for the MrNV CP degradation. The addition of E-64, a cysteine protease inhibitor, remarkably improved the yield of MrNV CP by 2.3-fold compared to the control. This innovative approach generates an economical method to improve the scalability of MrNV CP VLPs using individual protease inhibitors, enabling the protein to retain their structural integrity and stability for prominent downstream applications including drug delivery and vaccine development |
| first_indexed | 2025-11-14T21:18:41Z |
| format | Article |
| id | sunway-1835 |
| institution | Sunway University |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T21:18:41Z |
| publishDate | 2021 |
| publisher | MDPI |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | sunway-18352021-08-23T06:54:34Z http://eprints.sunway.edu.my/1835/ Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein Bethilda, A. S. Abdul Razak, M. Ho, K. L. Ng, C. L. Yong, Chean Yeah * Tan, W. S. QR355 Virology The causative agent of white tail disease (WTD) in the giant freshwater prawn is Macrobrachium rosenbergii nodavirus (MrNV). The recombinant capsid protein (CP) of MrNV was previously expressed in Escherichia coli, and it self-assembled into icosahedral virus-like particles (VLPs) with a diameter of approximately 30 nm. Extensive studies on the MrNV CP VLPs have attracted widespread attention in their potential applications as biological nano-containers for targeted drug delivery and antigen display scaffolds for vaccine developments. Despite their advantageous features, the recombinant MrNV CP VLPs produced in E. coli are seriously affected by protease degradations, which significantly affect the yield and stability of the VLPs. Therefore, the aim of this study is to enhance the stability of MrNV CP by modulating the protease degradation activity. Edman degradation amino acid sequencing revealed that the proteolytic cleavage occurred at arginine 26 of the MrNV CP. The potential proteases responsible for the degradation were predicted in silico using the Peptidecutter, Expasy. To circumvent proteolysis, specific protease inhibitors (PMSF, AEBSF and E-64) were tested to reduce the degradation rates. Modulation of proteolytic activity demonstrated that a cysteine protease was responsible for the MrNV CP degradation. The addition of E-64, a cysteine protease inhibitor, remarkably improved the yield of MrNV CP by 2.3-fold compared to the control. This innovative approach generates an economical method to improve the scalability of MrNV CP VLPs using individual protease inhibitors, enabling the protein to retain their structural integrity and stability for prominent downstream applications including drug delivery and vaccine development MDPI 2021-08 Article PeerReviewed text en cc_by_nc_nd_4 http://eprints.sunway.edu.my/1835/1/Yong%20Chean%20Yeah%20Regulation%20of%20proteolytic%20activity%20to%20improve%20the-ijms-22-08725.pdf Bethilda, A. S. and Abdul Razak, M. and Ho, K. L. and Ng, C. L. and Yong, Chean Yeah * and Tan, W. S. (2021) Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein. International Journal of Molecular Sciences, 22 (16). p. 8725. ISSN 1422-0067 http://doi.org/10.3390/ijms22168725 https://www.mdpi.com/1422-0067/22/16/8725 |
| spellingShingle | QR355 Virology Bethilda, A. S. Abdul Razak, M. Ho, K. L. Ng, C. L. Yong, Chean Yeah * Tan, W. S. Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title | Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title_full | Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title_fullStr | Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title_full_unstemmed | Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title_short | Regulation of Proteolytic activity to improve the recovery of Macrobrachium rosenbergii Nodavirus capsid protein |
| title_sort | regulation of proteolytic activity to improve the recovery of macrobrachium rosenbergii nodavirus capsid protein |
| topic | QR355 Virology |
| url | http://eprints.sunway.edu.my/1835/ http://eprints.sunway.edu.my/1835/ http://eprints.sunway.edu.my/1835/ http://eprints.sunway.edu.my/1835/1/Yong%20Chean%20Yeah%20Regulation%20of%20proteolytic%20activity%20to%20improve%20the-ijms-22-08725.pdf |