Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase
Abstract Disulfide bond engineering is a promising strategy for enhancing the stability and functional lifespan of enzymes in therapeutic and industrial applications. In this study, we applied computational modeling to introduce interchain disulfide bonds in Aspergillus flavus uricase to increase it...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-05-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-01683-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850231519985008640 |
|---|---|
| author | Mohammad Reza Rahbar Navid Nezafat Mohammad Hossein Morowvat Amir Savardashtaki Mohammad Bagher Ghoshoon Mohammad Soroosh Hajizade Younes Ghasemi |
| author_facet | Mohammad Reza Rahbar Navid Nezafat Mohammad Hossein Morowvat Amir Savardashtaki Mohammad Bagher Ghoshoon Mohammad Soroosh Hajizade Younes Ghasemi |
| author_sort | Mohammad Reza Rahbar |
| collection | DOAJ |
| description | Abstract Disulfide bond engineering is a promising strategy for enhancing the stability and functional lifespan of enzymes in therapeutic and industrial applications. In this study, we applied computational modeling to introduce interchain disulfide bonds in Aspergillus flavus uricase to increase its stability without compromising catalytic efficiency. Six uricase muteins were engineered with targeted disulfide bonds at positions selected based on energetic frustration, structural integrity, and tunnel profiling analyses. By employing frustration density mapping, Root Mean Square Fluctuation (RMSF) profiling, and tunnel analysis, we evaluated the structural stability, flexibility, and substrate accessibility of each variant. Our findings revealed that muteins with disulfide bonds between residues such as Ala6-Cys290 and Ser119-Cys220 exhibited significant reductions in highly frustrated regions, enhancing the enzyme’s structural resilience. RMSF analysis indicated decreased local flexibility near disulfide sites, contributing to increased stability. Tunnel profiling further demonstrated that muteins with strategically placed disulfide bonds maintained favorable substrate access and low-energy barriers, critical for catalytic turnover. These results underscore the potential of targeted disulfide bond engineering for optimizing enzyme stability, offering valuable insights for the development of stable, high-performance biocatalysts suitable for therapeutic and industrial use. |
| format | Article |
| id | doaj-art-0656af8d77264e6ebcae383b431e1126 |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-0656af8d77264e6ebcae383b431e11262025-08-20T02:03:31ZengNature PortfolioScientific Reports2045-23222025-05-0115112910.1038/s41598-025-01683-yStability and functional consequences of disulfide bond engineering in Aspergillus flavus uricaseMohammad Reza Rahbar0Navid Nezafat1Mohammad Hossein Morowvat2Amir Savardashtaki3Mohammad Bagher Ghoshoon4Mohammad Soroosh Hajizade5Younes Ghasemi6Pharmaceutical Sciences Research Center, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical SciencesDepartment of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical SciencesDepartment of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical SciencesPharmaceutical Sciences Research Center, Shiraz University of Medical SciencesAbstract Disulfide bond engineering is a promising strategy for enhancing the stability and functional lifespan of enzymes in therapeutic and industrial applications. In this study, we applied computational modeling to introduce interchain disulfide bonds in Aspergillus flavus uricase to increase its stability without compromising catalytic efficiency. Six uricase muteins were engineered with targeted disulfide bonds at positions selected based on energetic frustration, structural integrity, and tunnel profiling analyses. By employing frustration density mapping, Root Mean Square Fluctuation (RMSF) profiling, and tunnel analysis, we evaluated the structural stability, flexibility, and substrate accessibility of each variant. Our findings revealed that muteins with disulfide bonds between residues such as Ala6-Cys290 and Ser119-Cys220 exhibited significant reductions in highly frustrated regions, enhancing the enzyme’s structural resilience. RMSF analysis indicated decreased local flexibility near disulfide sites, contributing to increased stability. Tunnel profiling further demonstrated that muteins with strategically placed disulfide bonds maintained favorable substrate access and low-energy barriers, critical for catalytic turnover. These results underscore the potential of targeted disulfide bond engineering for optimizing enzyme stability, offering valuable insights for the development of stable, high-performance biocatalysts suitable for therapeutic and industrial use.https://doi.org/10.1038/s41598-025-01683-yDisulfide Bond EngineeringUricase StabilityEnergetic FrustrationProtein Folding DynamicsEnzyme Tunnel ProfilingIn Silico Protein Design |
| spellingShingle | Mohammad Reza Rahbar Navid Nezafat Mohammad Hossein Morowvat Amir Savardashtaki Mohammad Bagher Ghoshoon Mohammad Soroosh Hajizade Younes Ghasemi Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase Scientific Reports Disulfide Bond Engineering Uricase Stability Energetic Frustration Protein Folding Dynamics Enzyme Tunnel Profiling In Silico Protein Design |
| title | Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase |
| title_full | Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase |
| title_fullStr | Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase |
| title_full_unstemmed | Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase |
| title_short | Stability and functional consequences of disulfide bond engineering in Aspergillus flavus uricase |
| title_sort | stability and functional consequences of disulfide bond engineering in aspergillus flavus uricase |
| topic | Disulfide Bond Engineering Uricase Stability Energetic Frustration Protein Folding Dynamics Enzyme Tunnel Profiling In Silico Protein Design |
| url | https://doi.org/10.1038/s41598-025-01683-y |
| work_keys_str_mv | AT mohammadrezarahbar stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT navidnezafat stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT mohammadhosseinmorowvat stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT amirsavardashtaki stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT mohammadbagherghoshoon stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT mohammadsorooshhajizade stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase AT younesghasemi stabilityandfunctionalconsequencesofdisulfidebondengineeringinaspergillusflavusuricase |