Exploring the impact of the recombinant Escherichia coli strain on defensins antimicrobial activity: BL21 versus Origami strain
The growing emergence of microorganisms resistant to antibiotics has prompted the development of alternative antimicrobial therapies. Among them, the antimicrobial peptides produced by innate immunity, which are also known as host defense peptides (HDPs), hold great potential. They have been shown to exert activity against both Gram-positive and Gram-negative bacteria, including those resistant to antibiotics. These HDPs are classified into three categories: defensins, cathelicidins, and histatins. Traditionally, HDPs have been chemically synthesized, but this strategy often limits their application due to the high associated production costs. Alternatively, some HDPs have been recombinantly produced, but little is known about the impact of the bacterial strain in the recombinant product. This work aimed to assess the influence of the Escherichia coli strain used as cell factory to determine the activity and stability of recombinant defensins, which have 3 disulfide bonds. For that, an α-defensin [human α-defensin 5 (HD5)] and a β-defensin [bovine lingual antimicrobial peptide (LAP)] were produced in two recombinant backgrounds. The first one was an E. coli BL21 strain, which has a reducing cytoplasm, whereas the second was an E. coli Origami B, that is a strain with a more oxidizing cytoplasm. The results showed that both HD5 and LAP, fused to Green Fluorescent Protein (GFP), were successfully produced in both BL21 and Origami B strains. However, differences were observed in the HDP production yield and bactericidal activity, especially for the HD5-based protein. The HD5 protein fused to GFP was not only produced at higher yields in the E. coli BL21 strain, but it also showed a higher quality and stability than that produced in the Origami B strain. Hence, this data showed that the strain had a clear impact on both HDPs quantity and quality.
579 - Microbiology
Is part of
Microbial Cell Factories
López-Cano, Adrià, Marc Martínez-Miguel, Judith Guasch, Imma Ratera, Anna Arís, and Elena Garcia-Fruitós. 2022. "Exploring The Impact Of The Recombinant Escherichia Coli Strain On Defensins Antimicrobial Activity: BL21 Versus Origami Strain". Microbial Cell Factories 21 (1). doi:10.1186/s12934-022-01803-7.
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/
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/PID2019-105622RB-I00/ES/PROCESAMIENTO DE MOLECULAS PARA CREAR MATERIALES HIBRIDOS ESTRUCTURADOS JERARQUICAMENTE PARA APLICACIONES BIOMEDICAS/
MINECO/Programa Estatal de promoción del talento y su empleabilidad en I+D+I/RYC-2017-22614/ES/ /
MINECO/Programa Estatal de promoción del talento y su empleabilidad en I+D+I/SEV-2017-0706/ES/ /
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