Functional inclusion bodies produced in the yeast Pichia pastoris
Visualitza/Obre
Autor/a
Rueda, Fabián
Gasser, Brigitte
Sánchez‑Chardi, Alejandro
Roldán, Mònica
Villegas, Sandra
Puxbaum, Verena
Ferrer‑Miralles, Neus
Unzueta, Ugutz
Vázquez, Esther
Garcia‑Fruitós, Elena
Mattanovich, Diethard
Villaverde, Antonio
Data de publicació
2016-10-01ISSN
1475-2859
Resum
Background: Bacterial inclusion bodies (IBs) are non‑toxic protein aggregates commonly produced in recombinant bacteria. They are formed by a mixture of highly stable amyloid‑like fibrils and releasable protein species with a significant extent of secondary structure, and are often functional. As nano structured materials, they are gaining biomedical interest because of the combination of submicron size, mechanical stability and biological activity, together with their ability to interact with mammalian cell membranes for subsequent cell penetration in absence of toxicity. Since essentially any protein species can be obtained as IBs, these entities, as well as related protein clusters (e.g., aggresomes), are being explored in biocatalysis and in biomedicine as mechanically stable sources of functional protein. One of the major bottlenecks for uses of IBs in biological interfaces is their potential contamination with endotoxins from producing bacteria.
Results: To overcome this hurdle, we have explored here the controlled production of functional IBs in the yeast Pichia pastoris (Komagataella spp.), an endotoxin‑free host system for recombinant protein production, and determined the main physicochemical and biological traits of these materials. Quantitative and qualitative approaches clearly indicate the formation of IBs inside yeast, similar in morphology, size and biological activity to those produced in E. coli, that once purified, interact with mammalian cell membranes and penetrate cultured mammalian cells in absence of toxicity.
Conclusions: Structurally and functionally similar from those produced in E. coli, the controlled production of IBs in P.
pastoris demonstrates that yeasts can be used as convenient platforms for the biological fabrication of self‑organizing
protein materials in absence of potential endotoxin contamination and with additional advantages regarding, among
others, post‑translational modifications often required for protein functionality.
Tipus de document
Article
Versió del document
Versió publicada
Llengua
English
Matèries (CDU)
636 - Explotació i cria d'animals. Cria del bestiar i d'animals domèstics
Pàgines
12
Publicat per
BMC
Publicat a
Microbial Cell Factories
Citació
Rueda, Fabián, Brigitte Gasser, Alejandro Sánchez-Chardi, Mònica Roldán, Sandra Villegas, Verena Puxbaum, and Neus Ferrer-Miralles et al. 2016. "Functional Inclusion Bodies Produced In The Yeast Pichia Pastoris". Microbial Cell Factories 15 (1). doi:10.1186/s12934-016-0565-9.
Programa
Producció de Remugants
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