Accès ouvert

Novel non-specific lipid-Transfer Protein (TdLTP4) isolated from Durum wheat: Antimicrobial activities to control pathogen and spoilage bacteria and Anti-Inflammatory properties in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages

Article scientifique 2020 Anglais

Résumé

Abstract Background: Lipid transfer proteins (LTP) are members of the family of pathogenesis-related proteins (PR-14) that play a key role in plant defense mechanisms.Methods: In this study, a novel gene TdLTP4 encoding an antifungal protein from wheat (cv. Om Rabiaa) was subcloned, overexpressed in Escherichia coli BL-21 (DE3) and enriched using ammonium sulfate fractionation. The TdLTP4 fusion protein was then tested against a panel of pathogens, food-borne and spoilage bacteria and fungi in order to evaluate the antimicrobial properties. Our protein was applied to 0.5 µg/mL LPS-induced RAW 264.7 macrophages in vitro at different concentrations (5, 10, 20, 50 and 100 µg/ml). Levels of nitric oxide (NO), pro-inflammatory cytokines interleukin (IL)-1β (IL-1 β), interleukin (IL)-6 (IL-6), tumor necrosis factor (TNF-α) and anti-inflammatory cytokine IL-10 in the supernatant fraction were measured using enzyme-linked immunosorbent assay (ELISA). Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected via Western blot. Results: The inhibition zones and minimal inhibitory concentration (MIC) values of bacterial strains were in the range of 14-26 mm and 62.5-250 µg/mL, respectively. Moreover, a remarkable activity against several fungal strains was revealed. TdLTP4 (5–100 µg/mL) decreased the production of NO (IC50= 4.32 µg/mL), IL-6 (IC50= 11.52 µg/mL), IL-1β (IC50= 7.87 µg/mL) and TNF-α (IC50 =8.66 µg/mL) by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Our protein could modulate the macrophages inflammatory mode by causing reduction in iNOS and COX2.Conclusion: According to these findings, LTP fusion protein could be used as natural anti-inflammatory and antimicrobial agent in food preservation and human health.

Citer ce document

Hsouna, A., Saad, R., Dhifi, W., Khaled, M., Mnif, W., Brini, F. (2020). Novel non-specific lipid-Transfer Protein (TdLTP4) isolated from Durum wheat: Antimicrobial activities to control pathogen and spoilage bacteria and Anti-Inflammatory properties in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. https://doi.org/10.21203/rs.3.rs-37982/v1

Accès au document

Voir sur le dépôt source

Ce document est hébergé sur son dépôt institutionnel d'origine.

Auteur(s)

Anis Ben Hsouna Rania Ben Saad Wissal Dhifi Marwa Khaled Wissem Mnif Faïçal Brini

Statistiques

Consultations : 1

Téléchargements : 0