Proteomic analysis of the probiotic Lactobacillus reuteri CRL 1098 reveals novel tolerance biomarkers to bile acid-induced stress

Authors
Bustos, Ana Yanina; Font, Graciela Maria; Raya, Raul Ricardo; de Almeida, André Martinho; Fadda, Silvina G.; Taranto, Maria Pia
Publication Year
2015
Language
English
Format
article
Status
Published version
Description
Lactobacillus (L.) reuteri CRL1098 is a probiotic bacterium with a proven hypocholesterolemic effect, moderateimmune stimulant effect and ability to produce cobalamin. The CRL1098 strain survives the passage throughthe gastrointestinal tract where the exposure to bile acids (BA) causes deleterious effects. In order to characterizethe molecular mechanisms through which L. reuteri CRL1098 adapts to bile, its proteomic response was evaluatedin the presence of conjugated (glycodeoxycholic acid-GDCA-) and free (deoxycholic acid-DCA-) bile acids (BA).Cell growth inhibition was observed only in the presence of DCA. Two-dimensional gel electrophoresis coupled tomass spectrometry allowed us to identify 25 protein spots differentially expressed in response to both BA. The mainfunctional categories assigned to the proteins were metabolism of nucleotides and glycerolipids, transcription andtranslation, pH homeostasis and stress-responses. Remarkably, cytosine triphosphate(CTP) synthetase, enzymerelated to the repair of oxidative DNA, was over-expressed in the presence of GDCA and significantly repressed byDCA; also three proteins related to protein transcription and translation were over expressed in the presence ofthe conjugated BA and one, was repressed by the free BA. This differential expression could explain the delayedgrowth of the cells challenged with the free BA and the unaffected growth in the presence of GDCA. Moreover,some general stress proteins were triggered in the presence of both BA. In addition, the bile salt hydrolase (BSH)enzyme regulation in response to BA was analyzed using real time-PCR to determine its contribution to cell tolerance. An up-regulation of the bsh gene in response to BA was observed, suggesting that this enzyme could be a specific biomarker of bile adaption in L. reuteri CRL1098. The present work proposes that BA induce a complexphysiological response in L. reuteri and provide new insights into the mechanisms involved in BA tolerance.
Fil: Bustos, Ana Yanina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Font, Graciela Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina
Fil: Raya, Raul Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: de Almeida, André Martinho. Centro Interdisciplinar de Investigação Em Sanidade Ani; Portugal. Universidade Nova de Lisboa. Instituto de Tecnologia Quimica e Biológica; Portugal. Instituto de Biologia Experimental e Tecnológica; Portugal
Fil: Fadda, Silvina G.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Taranto, Maria Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Subject
LACTOBACILLUS REUTERI
BILE ACID TOLERANCE
TWO-DIMENSIONAL ELECTROPHORESIS
BILE SALT HYDROLASE
REAL TIME-PCR
Otras Ciencias Biológicas
Ciencias Biológicas
CIENCIAS NATURALES Y EXACTAS
Access level
Restricted access
License
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/28459