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1.
Nutrire Rev. Soc. Bras. Aliment. Nutr ; 41: 1-10, Dec. 2016. tab, ilus
Artículo en Inglés | LILACS | ID: biblio-880608

RESUMEN

BACKGROUND: Eight Lactobacillus reuteri strains, previously isolated from breast-fed human infant feces, were selected to assess the potential contribution of their surface proteins in probiotic activity. These strains were treated with 5 M LiCl to remove their surface proteins, and their tolerance to simulated stomach-duodenum passage, cell surface characteristics, auto aggregation, adhesion, and inhibition of pathogen adhesion to Caco-2 cells were compared with untreated strains. RESULTS: The survival rates, auto aggregation, and adhesion abilities of the LiCl-treated L. Reuteri strains decreased significantly (p< 0.05) compared to that of the untreated cells. The inhibition ability of selected L. reuteri strains, untreated or LiCl treated, against adherence of Escherichia coli 25922 and Salmonella typh iNCDC113 to Caco-2 was evaluated in vitro with L. reuteri ATCC55730 strain as a positive control. Among the selected eight strains of L.reuteri, LR6 showed maximum inhibition against the E. Coli ATCC25922 and S. typhiNCDC113. After treatment with 5 M LiCl to remove surface protein, the inhibition activities of the lactobacilli against pathogens decreased significantly (p< 0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated thatLR6 strains had several bands with molecular weight ranging from 10 to 100 KDa, and their characterization and functions need to be confirmed. CONCLUSIONS: The results revealed that the cell surface proteins of L. reuteri play an important role in their survivability, adhesion, and competitive exclusion of pathogen to epithelial cells.


Asunto(s)
Limosilactobacillus reuteri/química , Limosilactobacillus reuteri/inmunología , Proteínas de la Membrana/metabolismo , Probióticos/uso terapéutico
2.
JMB-Journal of Medical Bacteriology. 2012; 1 (3,4): 10-16
en Inglés | IMEMR | ID: emr-139761

RESUMEN

The prevalence of Urinary Tract Infection [UTI] is really high in the world. Escherichia coli is a major agent of UTI. One of the strategies for decreasing UTI infections is vaccine development. As the attachment is a really important stage in colonization and infection, attachment inhibition has an applied strategy. FimH protein is a major factor during bacterial colonization in urinary tract and could be used as a vaccine. Thus, it was considered in this research as a candidate antigen. The sequences of fimH and acmA genes were used for designing a synthetic gene. It was cloned to pET23a expression vector and transformed to E. coli [DE3] Origami. To confirm the expression of recombinant protein, SDS-PAGE and western blotting methods were used. Subsequently, recombinant protein was purified. On the other hand, Lactobacillus reuteri was cultured and mixed with FimH / AcmA recombinant protein. The rate of protein localization on lactobacillus surface was assessed using ELISA method. It was showed that the recombinant protein was expressed in E. coli [DE3] Origami and purified by affinity chromatography. Moreover, this protein could be localized on lactobacillus surface by 5 days. In current study, a fusion recombinant protein was prepared and displayed on L. reuteri surface. This strain could be used for animal experiment as a competitor against Uropathogenic E. coli [UPEC]. Using manipulated probiotics strains instead of antibiotic therapy could decrease the antibiotic consumption and reduce multi-drug resistant strains


Asunto(s)
Limosilactobacillus reuteri/genética , Limosilactobacillus reuteri/inmunología , Infecciones Urinarias/genética , Ensayo de Inmunoadsorción Enzimática , Antiinfecciosos Urinarios , Probióticos , Escherichia coli/genética
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