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Microbial Cell Factories

, 9:71

First Online: 17 September 2010Received: 08 May 2010Accepted: 17 September 2010

Abstract

BackgroundBacterial inclusion bodies are submicron protein clusters usually found in recombinant bacteria that have been traditionally considered as undesirable products from protein production processes. However, being fully biocompatible, they have been recently characterized as nanoparticulate inert materials useful as scaffolds for tissue engineering, with potentially wider applicability in biomedicine and material sciences. Current protocols for inclusion body isolation from Escherichia coli usually offer between 95 to 99% of protein recovery, what in practical terms, might imply extensive bacterial cell contamination, not compatible with the use of inclusion bodies in biological interfaces.

ResultsUsing an appropriate combination of chemical and mechanical cell disruption methods we have established a convenient procedure for the recovery of bacterial inclusion bodies with undetectable levels of viable cell contamination, below 10 cfu-ml, keeping the particulate organization of these aggregates regarding size and protein folding features.

ConclusionsThe application of the developed protocol allows obtaining bacterial free inclusion bodies suitable for use in mammalian cell cultures and other biological interfaces.

Electronic supplementary materialThe online version of this article doi:10.1186-1475-2859-9-71 contains supplementary material, which is available to authorized users.

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Autor: Escarlata Rodríguez-Carmona - Olivia Cano-Garrido - Joaquin Seras-Franzoso - Antonio Villaverde - Elena García-Fruitós

Fuente: https://link.springer.com/







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