Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/45777

TitleAntibiotic free selection for the high level biosynthesis of a silk-elastin-like protein
Author(s)Barroca, Mário Jorge Faria
Rodrigues, Paulo
Sobral, Rómulo Sacramento
Costa, Maria Manuela Ribeiro
Chaves, Susana R.
Machado, Raul
Casal, Margarida
Collins, Tony
Issue date16-Dec-2016
PublisherNature Publishing Group
JournalScientific Reports
CitationBarroca M., Rodrigues P., Sobral R., Costa M.M.R., Chaves S.R., Machado R., Casal M. and Collins T. Antibiotic free selection for the high level biosynthesis of a silk-elastin-like protein. (2016). Scientific Reports, 6, 39329; DOI: 10.1038/srep39329.
Abstract(s)Silk-elastin-like proteins (SELPs) are a family of genetically engineered recombinant protein polymers exhibiting mechanical and biological properties suited for a wide range of applications in the biomedicine and materials fields. They are being explored as the next generation of biomaterials but low productivities and use of antibiotics during production undermine their economic viability and safety. We have developed an industrially relevant, scalable, fed-batch process for the high level production of a novel SELP in E. coli in which the commonly used antibiotic selection marker of the expression vector is exchanged for a post segregational suicide system, the separate-component-stabilisation system (SCS). SCS significantly augments SELP productivity but also enhances the product safety profile and reduces process costs by eliminating the use of antibiotics. Plasmid content increased following induction but no significant differences in plasmid levels were discerned when using SCS or the antibiotic selection markers under the controlled fed-batch conditions employed. It is suggested that the absence of competing plasmid-free cells improves host cell viability and enables increased productivity with SCS. With the process developed, 12.8 g L(-1) purified SELP was obtained, this is the highest SELP productivity reported to date and clearly demonstrates the commercial viability of these promising polymers.
TypeArticle
URIhttp://hdl.handle.net/1822/45777
DOI10.1038/srep39329
ISSN2045-2322
Publisher versionhttps://www.nature.com/articles/srep39329
Peer-Reviewedyes
AccessOpen access
Appears in Collections:DBio - Artigos/Papers

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