Time to consider ruling out inclusion bodies denaturing protocols for spontaneous solubilization of biologically active proteins.
Activity assay
Inclusion bodies
Recombinant production
Solubilization
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
30 10 2024
30 10 2024
Historique:
received:
04
03
2024
accepted:
25
10
2024
medline:
30
10
2024
pubmed:
30
10
2024
entrez:
30
10
2024
Statut:
epublish
Résumé
The formation of inclusion bodies (IBs) in microbial cell factories is a very common process occurring during recombinant protein production. Different protocols have been developed for the extraction of soluble proteins from IBs using several strategies, ranging from the use of harsh denaturing and high concentrations of chaotropic agents and reducing agents to the use of mild protocols based on the use of non-denaturing detergents. However, in recent years, the biological vision of IBs has changed and research studies have demonstrated that these protein aggregates contain biologically active and properly folded recombinant proteins. This drives us to redefine the methodologies currently used to obtain soluble protein using IB as a protein source. Hence, we propose the extraction of IB protein via the simple spontaneous solubilization of IB as a strategy broadly applicable to all kinds of recombinant proteins without the negative effects of detergents and chaotropic agents on final biological activity. We prove the wide applicability of spontaneous solubilization processes to different types of IBs and that protocols can be easily customized for each protein in terms of timing and incubation temperature by monitoring the protein activity of the solubilized fraction.
Identifiants
pubmed: 39472505
doi: 10.1038/s41598-024-77899-1
pii: 10.1038/s41598-024-77899-1
doi:
Substances chimiques
Recombinant Proteins
0
Detergents
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
26061Subventions
Organisme : Ministerio de Ciencia, Innovación y Universidades
ID : Grants PID2019-107298RB-C21/AEI/10.13039/ 501100011033
Organisme : Ministerio de Ciencia, Innovación y Universidades
ID : Grants PID2019-107298RB-C21/AEI/10.13039/ 501100011033
Organisme : Ministerio de Ciencia, Innovación y Universidades
ID : Grants PID2019-107298RB-C21/AEI/10.13039/ 501100011033
Organisme : AGAUR
ID : 2021 SGR 01552.
Organisme : AGAUR
ID : 2021 SGR 01552.
Informations de copyright
© 2024. The Author(s).
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