Exploring the impact of the recombinant Escherichia coli strain on defensins antimicrobial activity : BL21 versus Origami strain
López-Cano, Adrià (Universitat Autònoma de Barcelona. Unitat Mixta d'Investigació IRTA-UAB en Sanitat Animal)
Martínez-Miguel, Marc (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)
Guasch, Judith (Max Planck Partner Group. Dynamic Biomimetics for Cancer Immunotherapy)
Ratera Bastardas, Imma (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)
Arís i Giralt, Anna (Universitat Autònoma de Barcelona. Unitat Mixta d'Investigació IRTA-UAB en Sanitat Animal)
Garcia-Fruitos, Elena (Universitat Autònoma de Barcelona. Unitat Mixta d'Investigació IRTA-UAB en Sanitat Animal)

Fecha:	2022
Resumen:	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.
Ayudas:	Agencia Estatal de Investigación PID2019-105622RB-I00 Agencia Estatal de Investigación PID2019-107298RB-C21 Ministerio de Ciencia e Innovación CEX2019-000917-S Fundació la Marató de TV3 201812-30-31-32-33
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Host defense peptides ; Escherichia coli ; Strain ; Recombinant protein
Publicado en:	Microbial cell factories, Vol. 21 (may 2022) , ISSN 1475-2859

DOI: 10.1186/s12934-022-01803-7
PMID: 35527241

10 p, 1.3 MB

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Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias de la salud y biociencias > Centre de Recerca en Sanitat Animal (CReSA-IRTA)
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