Biochemical methods for producing and characterising recombinant spider silks
Here, we review the processes involved in producing and assessing the quality of recombinant spider silk proteins (spidroins) and the challenges associated with their synthesis and spinning into robust fibres. We provide an overview of the techniques used to produce the proteins, from gene synthesis...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Arachnid Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/frchs.2024.1488680/full |
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| author | Anton Maraldo Anton Maraldo James H. Torpey Sasha L. Evans Aditya Rawal Jeffery L. Yarger Rivka L. Isaacson Christopher P. Marquis Christopher P. Marquis Sean J. Blamires Sean J. Blamires |
| author_facet | Anton Maraldo Anton Maraldo James H. Torpey Sasha L. Evans Aditya Rawal Jeffery L. Yarger Rivka L. Isaacson Christopher P. Marquis Christopher P. Marquis Sean J. Blamires Sean J. Blamires |
| author_sort | Anton Maraldo |
| collection | DOAJ |
| description | Here, we review the processes involved in producing and assessing the quality of recombinant spider silk proteins (spidroins) and the challenges associated with their synthesis and spinning into robust fibres. We provide an overview of the techniques used to produce the proteins, from gene synthesis to expression in various host organisms. Evidence suggests that the N- and C-terminal regions of spidroins are of utmost importance for fibre assembly and the repetitive domains are responsible for the unique mechanical properties in both native and recombinant versions of spider silks. We describe the role of liquid–liquid phase separation (LLPS) in spidroin assembly and its importance in subsequent fibre formation. Recent developments in recombinant spidroin production and co-expression strategies for improving yield and scalability are highlighted. Techniques such as mass photometry and size exclusion chromatography (SEC) for analysing protein purity and assembly behaviour are thereupon detailed. Finally, we address the role that predictive computational methods play in the future of designing novel and high-performing materials inspired by spidroins. |
| format | Article |
| id | doaj-art-8b76f998b5724b0693aa3a9dd3b27cea |
| institution | Kabale University |
| issn | 2813-5083 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Arachnid Science |
| spelling | doaj-art-8b76f998b5724b0693aa3a9dd3b27cea2025-01-21T08:36:35ZengFrontiers Media S.A.Frontiers in Arachnid Science2813-50832025-01-01310.3389/frchs.2024.14886801488680Biochemical methods for producing and characterising recombinant spider silksAnton Maraldo0Anton Maraldo1James H. Torpey2Sasha L. Evans3Aditya Rawal4Jeffery L. Yarger5Rivka L. Isaacson6Christopher P. Marquis7Christopher P. Marquis8Sean J. Blamires9Sean J. Blamires10School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, AustraliaRecombinant Products Facility, University of New South Wales, Sydney, NSW, AustraliaDepartment of Chemistry, King’s College London, London, United KingdomDepartment of Chemistry, King’s College London, London, United KingdomNuclear Magnetic Resonance (NMR) Facility, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, NSW, AustraliaSchool of Molecular Sciences, Arizona State University, Tempe, AZ, United StatesDepartment of Chemistry, King’s College London, London, United KingdomSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, AustraliaRecombinant Products Facility, University of New South Wales, Sydney, NSW, AustraliaDepartment of Chemistry, King’s College London, London, United KingdomSchool of Biological Earth and Environmental Sciences, University of New South Wales, Sydney, NSW, AustraliaHere, we review the processes involved in producing and assessing the quality of recombinant spider silk proteins (spidroins) and the challenges associated with their synthesis and spinning into robust fibres. We provide an overview of the techniques used to produce the proteins, from gene synthesis to expression in various host organisms. Evidence suggests that the N- and C-terminal regions of spidroins are of utmost importance for fibre assembly and the repetitive domains are responsible for the unique mechanical properties in both native and recombinant versions of spider silks. We describe the role of liquid–liquid phase separation (LLPS) in spidroin assembly and its importance in subsequent fibre formation. Recent developments in recombinant spidroin production and co-expression strategies for improving yield and scalability are highlighted. Techniques such as mass photometry and size exclusion chromatography (SEC) for analysing protein purity and assembly behaviour are thereupon detailed. Finally, we address the role that predictive computational methods play in the future of designing novel and high-performing materials inspired by spidroins.https://www.frontiersin.org/articles/10.3389/frchs.2024.1488680/fullrecombinant spider silkspidroin synthesisfibre assemblyliquid–liquid phase separationpredictive computational model |
| spellingShingle | Anton Maraldo Anton Maraldo James H. Torpey Sasha L. Evans Aditya Rawal Jeffery L. Yarger Rivka L. Isaacson Christopher P. Marquis Christopher P. Marquis Sean J. Blamires Sean J. Blamires Biochemical methods for producing and characterising recombinant spider silks Frontiers in Arachnid Science recombinant spider silk spidroin synthesis fibre assembly liquid–liquid phase separation predictive computational model |
| title | Biochemical methods for producing and characterising recombinant spider silks |
| title_full | Biochemical methods for producing and characterising recombinant spider silks |
| title_fullStr | Biochemical methods for producing and characterising recombinant spider silks |
| title_full_unstemmed | Biochemical methods for producing and characterising recombinant spider silks |
| title_short | Biochemical methods for producing and characterising recombinant spider silks |
| title_sort | biochemical methods for producing and characterising recombinant spider silks |
| topic | recombinant spider silk spidroin synthesis fibre assembly liquid–liquid phase separation predictive computational model |
| url | https://www.frontiersin.org/articles/10.3389/frchs.2024.1488680/full |
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