This study aimed to isolate Lactobacillus species from yoghurt and to evaluate its antimicrobial activity against enterohemorrhagic Escherichia coli O157:H7. After incubation of different yoghurt samples in MRS (de Man, Rogosa and Sharpe) agar at 37C under anaerobic conditions, four Lactobacillus species, L. casei, L. acidophilus, L. helveticus and L. delbrueckii ssp. bulgaricus were identified. Standard growth curves of E. coli O157:H7 in proximity with lactobacilli test strains were obtained. The antibacterial effects of lactobacilli cell debris and culture supernatant against E. coli O157:H7 were determined. The results showed that the antibacterial effects of culture supernatant were stable at different temperature conditions (56-100C) for 30 and 60 min; also, it was stable at pH 3-10. With regard to the antibacterial effect of the lactobacilli strains in this study, they may help in the treatment of E. coli O157:H7-associated diseases and can be applied as a biological preservative in the food industry.
Background and purpose:
Recently, the use of immunotoxins for targeted cancer therapy has been proposed, to find new anticancer drugs with high efficacy on tumor cells with minimal side effects on normal cells. we designed and compared several arazyme (AraA)-based fusion proteins with different ligands to choose the best-targeted therapy for interleukin 13 receptor alpha 2 (IL13Rα2)-overexpressed cancer cells. For this purpose, IL13Rα2 was selected as a receptor and IL13 and IL13.E13K were evaluated as native and mutant ligands, respectively. In addition, Pep-1 and A2b11 were chosen as the peptide ligands for targeted cancer therapy.
Experimental approach:
Several bioinformatics servers were used for designing constructs and optimization. The structures of the chimeric proteins were predicted and verified by I-TASSER, Q-Mean, ProSA, Ramachandran plot, and Verify3D program. Physicochemical properties, toxicity, and antigenicity were predicted by ProtParam, ToxinPred, and VaxiJen. HawkDock, LigPlot
+
, and GROMACS software were used for docking and molecular dynamics simulation of the ligand-receptor interaction.
Findings/Results:
The
in silico
results showed AraA-A2b11 has higher values of confidence score and Q-mean score was obtained for high-resolution crystal structures. All chimeric proteins were stable, non-toxic, and non-antigenic. AraA-(A(EAAAK)
4
ALEA(EAAAK)
4
A)
2
-IL13 retained its natural structure and based on ligand-receptor docking and molecular dynamic analysis, the binding ability of AraA-(A(EAAAK)
4
ALEA(EAAAK)
4
A)
2
-IL13 to IL13Rα2 was sufficiently strong.
Conclusion and implications:
Based on the bioinformatics result AraA-(A(EAAAK)
4
ALEA(EAAAK)
4
A)
2
-IL13 was a stable fusion protein with two separate domains and high affinity with the IL13Rα2 receptor. Therefore, AraA-(A(EAAAK)
4
ALEA(EAAAK)
4
A)
2
-IL13 fusion protein could be a new potent candidate for target cancer therapy.
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