Quality comparison of recombinant soluble proteins and proteins solubilized from bacterial inclusion bodies
Recombinant protein production in bacteria is often accompanied by the formation of aggregates, known as inclusion bodies (IBs). Although several strategies have been developed to minimize protein aggregation, many heterologous proteins are produced in aggregated form. For these proteins, purification necessarily requires processes of solubilization and refolding, often involving denaturing agents. However, the presence of biologically active recombinant proteins forming IBs has driven a redefinition of the protocols used to obtain soluble protein avoiding the protein denaturation step. Among the different strategies described, the detergent n-lauroylsarcosine (NLS) has proved to be effective. However, the impact of the NLS on final protein quality has not been evaluated so far. Here, the activity of three antimicrobial proteins (all as GFP fusions) obtained from the soluble fraction was compared with those solubilized from IBs. Results showed that NLS solubilized proteins from IBs efficiently, but that protein activity was impaired. Thus, a solubilization protocol without detergents was evaluated, demonstrating that this strategy efficiently solubilized proteins embedded in IBs while retaining their biological activity. These results showed that the protocol used for IB solubilization has an impact on final protein quality and that IBs can be solubilized through a very simple step, obtaining fully active proteins.
579 - Microbiology
Is part of
López-Cano, Adrià, Paula Sicilia, Clara Gaja, Anna Arís, and Elena Garcia-Fruitós. 2022. "Quality Comparison Of Recombinant Soluble Proteins And Proteins Solubilized From Bacterial Inclusion Bodies". New Biotechnology 72: 58-63. doi:10.1016/j.nbt.2022.09.003.
Grant agreement number
MICIU/Programa Estatal de generación del conocimiento y fortalecimiento científico y tecnológico del sistema I+D+I y Programa Estatal de I+D+I orientada a los retos de la sociedad/PID2019-107298RB-C21/ES/PRODUCCION Y VALIDACION DE FARMACOS BASADOS EN PEPTIDOS DE DEFENSA DEL HUESPED PARA EL TRATAMIENTO DEL SINDROME RESPIRATORIO BOVINO/
Producció de Remugants
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