Isolation of cell-free bacterial inclusion bodies

Microbial Cell Factories

Technical Notes

Elena García-Fruitós¹  Joaquin Seras-Franzoso²  Escarlata Rodríguez-Carmona²  Antonio Villaverde²  Olivia Cano-Garrido² 

[1] CIBER en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, 0819, Barcelona, Spain;Institut de Biotecnologia i de Biomedicina and Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), 0819, Barcelona, Spain;Institut de Biotecnologia i de Biomedicina and Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), 0819, Barcelona, Spain;CIBER en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, 0819, Barcelona, Spain;

关键词: Viable Bacterium;    Viable Cell Count;    Bacterial Lysis;    Lysozyme Treatment;    Sonication Cycle;   

DOI  :  10.1186/1475-2859-9-71

 received in 2010-05-08, accepted in 2010-09-17,  发布年份 2010

来源: Springer

【 摘 要 】

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 10-1 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.

【 授权许可】

Unknown   
© Rodríguez-Carmona et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.