Abstract

The production of pure and soluble proteins is a complex, protein-dependent and time-consuming process, in particular for those prone-to-aggregate and/or difficult-to-purify. Although Escherichia coli is widely used for protein production, recombinant products must be co-purified through costly processes to remove lipopolysaccharide (LPS) and minimize adverse effects in the target organism. Interestingly, Lactococcus lactis, which does not contain LPS, could be a promising alternative for the production of relevant proteins. However, to date, there is no universal strategy to produce and purify any recombinant protein, being still a protein-specific process. In this context and considering that L. lactis is also able to form functional protein aggregates under overproduction conditions, we explored the use of these aggregates as an alternative source of soluble proteins. In this study, we developed a widely applicable and economically affordable protocol to extract functional proteins from these nanoclusters. For that, two model proteins were used: mammary serum amyloid A3 (M-SAA3) and metalloproteinase 9 (MMP-9), a difficult-to-purify and a prone-to-aggregate protein, respectively. The results show that it is possible to obtain highly pure, soluble, LPS-free and active recombinant proteins from L. lactis aggregates through a cost-effective and simple protocol with special relevance for difficult-to-purify or highly aggregated proteins.

Details

Title
A new approach to obtain pure and active proteins from Lactococcus lactis protein aggregates
Author
Gifre-Renom, L 1 ; Cano-Garrido, O 2 ; Fàbregas, F 1 ; Roca-Pinilla, R 1 ; Seras-Franzoso, J 3 ; Ferrer-Miralles, N 2 ; Villaverde, A 2 ; Bach, À 4 ; Devant, M 1 ; Arís, A 1 ; Garcia-Fruitós, E 1   VIAFID ORCID Logo 

 Department of Ruminant Production, Institut de Recerca i Tecnologia Agroalimentàries (IRTA), Caldes de Montbui, Spain 
 Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Cerdanyola del Vallès, Spain 
 Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Cerdanyola del Vallès, Spain; Cibbim-Nanomedicine, Hospital Vall d’Hebron, Institut de Recerca de la Vall d’Hebron (VHIR), Barcelona, Spain 
 Department of Ruminant Production, Institut de Recerca i Tecnologia Agroalimentàries (IRTA), Caldes de Montbui, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain 
Pages
1-10
Publication year
2018
Publication date
Sep 2018
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2108211220
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.