scholarly journals Impact of luxS and Suppressor Mutations on the Gastrointestinal Transit of Lactobacillus rhamnosus GG

2008 ◽  
Vol 74 (15) ◽  
pp. 4711-4718 ◽  
Author(s):  
Sarah Lebeer ◽  
Ingmar J. J. Claes ◽  
Tine L. A. Verhoeven ◽  
Chong Shen ◽  
Ivo Lambrichts ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Gastric Juice ◽  
Biofilm Formation ◽  
Lactobacillus Rhamnosus ◽  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
Promoting Effect ◽  
Autoinducer 2 ◽  
Metabolic Defects ◽  
Suppressor Mutations

ABSTRACT It is generally believed that probiotic bacteria need to survive gastrointestinal transit to exert a health-promoting effect. In this study, a genuine luxS mutant and a luxS mutant containing unknown suppressor mutations of the probiotic strain Lactobacillus rhamnosus GG were compared to the wild type for survival and persistence in the murine gastrointestinal tract. The LuxS enzyme, catalyzing the production of the autoinducer-2 signaling molecule, also forms an integral part of the activated methyl cycle and the metabolism of methionine and cysteine. The genuine luxS mutant CMPG5412 showed drastically reduced persistence in mice, which was related to less survival in simulated gastric juice, indicating that LuxS metabolism is crucial for the gastric stress resistance of L. rhamnosus GG. The suppressor mutations in the other luxS mutant, CMPG5413, appear to compensate for the metabolic defects of the luxS mutation and to restore the resistance to gastric juice but cause a defect in adherence, biofilm formation, and exopolysaccharide production. The shorter residence time of this suppressor mutant in the murine gastrointestinal tract indicates a role for biofilm formation and exopolysaccharides in the persistence capacity of L. rhamnosus GG.

scholarly journals Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

2005 ◽  
Vol 71 (6) ◽  
pp. 3060-3067 ◽  
Author(s):  
B. M. Corcoran ◽  
C. Stanton ◽  
G. F. Fitzgerald ◽  
R. P. Ross
Keyword(s):  
Gastric Juice ◽  
Lactobacillus Rhamnosus ◽  
Probiotic Strain ◽  
Iodoacetic Acid ◽  
Glucose Effect ◽  
Ph Values ◽  
Gapdh Activity ◽  
Acidic Conditions ◽  
Probiotic Lactobacilli ◽  
Effect Of Glucose

ABSTRACT Lactobacillus rhamnosus GG is an industrially significant probiotic strain with proven health benefits. In this study, the effect of glucose on L. rhamnosus GG survival was analyzed in simulated gastric juice at pH 2.0. It was found that the presence of 19.4 mM glucose resulted in up to 6-log10-enhanced survival following 90 min of exposure. Further work with dilute HCl confirmed that glucose was the sole component responsible. Comparative analysis with other Lactobacillus strains revealed that enhanced survival was apparent in all strains, but at different pH values. The presence of glucose at concentrations from 1 to 19.4 mM enhanced L. rhamnosus GG survival from 6.4 to 8 log10 CFU ml−1 in simulated gastric juice. The mechanisms behind the protective effect of glucose were investigated. Addition of N′,N′-dicyclohexylcarbodiimide to simulated gastric juice caused survival to collapse, which was indicative of a prominent role in inhibition of F0F1-ATPase. Further work with neomycin-resistant mutants that exhibited 38% to 48% of the F0F1-ATPase activity of the parent confirmed this, as the survival in the presence of glucose of these mutants decreased 3 × 106-fold compared with the survival of the wild type (which had a viability of 8.02 log10 CFU ml−1). L. rhamnosus GG survival in acidic conditions occurred only in the presence of sugars that it could metabolize efficiently. To confirm the involvement of glycolysis in the glucose effect, iodoacetic acid was used to inhibit glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. The reduction in GAPDH activity caused survival to decrease by 8.30 log10 CFU ml−1 in the presence of glucose. The data indicate that glucose provides ATP to F0F1-ATPase via glycolysis, enabling proton exclusion and thereby enhancing survival during gastric transit.

scholarly journals Impact of Environmental and Genetic Factors on Biofilm Formation by the Probiotic Strain Lactobacillus rhamnosus GG

2007 ◽  
Vol 73 (21) ◽  
pp. 6768-6775 ◽  
Author(s):  
Sarah Lebeer ◽  
Tine L. A. Verhoeven ◽  
M�nica Perea V�lez ◽  
Jos Vanderleyden ◽  
Sigrid C. J. De Keersmaecker
Keyword(s):  
Biofilm Formation ◽  
Culture Medium ◽  
Lactobacillus Rhamnosus ◽  
Probiotic Strain ◽  
Lipoteichoic Acid ◽  
Microtiter Plate ◽  
Phenotypic Analysis ◽  
Lactobacillus Rhamnosus Gg ◽  
Abiotic Surfaces

ABSTRACTLactobacillus rhamnosusGG (ATCC 53103) is one of the clinically best-studied probiotic organisms. Moreover,L. rhamnosusGG displays very good in vitro adherence to epithelial cells and mucus. Here, we report thatL. rhamnosusGG is able to form biofilms on abiotic surfaces, in contrast to other strains of theLactobacillus caseigroup tested under the same conditions. Microtiter plate biofilm assays indicated that in vitro biofilm formation byL. rhamnosusGG is strongly modulated by culture medium factors and conditions related to the gastrointestinal environment, including low pH; high osmolarity; and the presence of bile, mucins, and nondigestible polysaccharides. Additionally, phenotypic analysis of mutants affected in exopolysaccharides (wzb), lipoteichoic acid (dltD), and central metabolism (luxS) showed their relative importance in biofilm formation byL. rhamnosusGG.

scholarly journals Functional Analysis of luxS in the Probiotic Strain Lactobacillus rhamnosus GG Reveals a Central Metabolic Role Important for Growth and Biofilm Formation

2006 ◽  
Vol 189 (3) ◽  
pp. 860-871 ◽  
Author(s):  
Sarah Lebeer ◽  
Sigrid C. J. De Keersmaecker ◽  
Tine L. A. Verhoeven ◽  
Abeer A. Fadda ◽  
Kathleen Marchal ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Lactobacillus Rhamnosus ◽  
In Silico Analysis ◽  
Probiotic Strain ◽  
Transcriptional Analysis ◽  
Knockout Mutant ◽  
Gastrointestinal Microbiota ◽  
Gene Encoding ◽  
Metabolic Role ◽  
Adaptive Processes

ABSTRACT Quorum sensing is involved in the regulation of multicellular behavior through communication via small molecules. Given the high number and diversity of the gastrointestinal microbiota, it is postulated that members of this community communicate to coordinate a variety of adaptive processes. AI-2 is suggested to be a universal bacterial signaling molecule synthesized by the LuxS enzyme, which forms an integral part of the activated methyl cycle. We have previously reported that the well-documented probiotic strain Lactobacillus rhamnosus GG, a human isolate, produces AI-2-like molecules. In this study, we identified the luxS homologue of L. rhamnosus GG. luxS seems to be located in an operon with a yxjH gene encoding a putative cobalamin-independent methionine synthase. In silico analysis revealed a methionine-specific T box in the leader sequence of the putative yxjH-luxS operon. However, transcriptional analysis showed that luxS is expressed mainly as a monocistronic transcript. Construction of a luxS knockout mutant confirmed that the luxS gene is responsible for AI-2 production in L. rhamnosus GG. However, this mutation also resulted in pleiotropic effects on the growth of this fastidious strain. Cysteine, pantothenate, folic acid, and biotin could partially complement growth, suggesting a central metabolic role for luxS in L. rhamnosus GG. Interestingly, the luxS mutant also showed a defect in monospecies biofilm formation. Experiments with chemically synthesized (S)-4,5-dihydroxy-2,3-pentanedione, coculture with the wild type, and nutritional complementation suggested that the main cause of this defect has a metabolic nature. Moreover, our data indicate that suppressor mutations are likely to occur in luxS mutants of L. rhamnosus GG. Therefore, results of luxS-related studies should be carefully interpreted.

In vitro probiotic characterization ofLactobacillusstrains from fermented radish and their anti-adherence activity against enteric pathogens

2015 ◽  
Vol 61 (11) ◽  
pp. 837-850 ◽  
Author(s):  
Karthiyaini Damodharan ◽  
Sasikumar Arunachalam Palaniyandi ◽  
Seung Hwan Yang ◽  
Joo-Won Suh
Keyword(s):  
Cell Viability ◽  
Lactobacillus Rhamnosus ◽  
Skim Milk ◽  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
Bile Salt Hydrolase ◽  
Probiotic Properties ◽  
Lactobacillus Pentosus ◽  
Shigella Boydii

In this study, we evaluated the probiotic properties of Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus fermentum strains isolated from fermented radish. All the strains survived the simulated oro-gastrointestinal transit condition and showed significantly higher adherence to Caco-2 cells compared with the probiotic strain Lactobacillus rhamnosus GG. The strains showed broad-spectrum antimicrobial activity, autoaggregation, and coaggregation capacity with pathogens. Furthermore, the Lactobacillus strains inhibited the adherence of Yersinia enterocolitica subsp. enterocolitica, Shigella boydii, and Salmonella choleraesuis to the Caco-2 cell line. The strains possessed bile salt hydrolase activity and their cholesterol-lowering activity in vitro was above 50% in the presence of bile. Strains of L. plantarum and L. pentosus possessed the plantaricin-encoding plnEF gene. In addition, the Lactobacillus strains maintained about 80% cell viability after freeze–drying in the presence of a combination of 5% skim milk and 5% maltodextrin as cryoprotectant, and 70% recovery of cell viability was observed in the absence of any cryoprotectant.

scholarly journals Preliminary evaluation of potential prebiotic capacity of selected legumes and seed mucilage on the probiotic strain Lactobacillus rhamnosus GG

2021 ◽  
pp. 60-72
Author(s):  
Ka Wai Lai ◽  
Yu Hsuan How ◽  
Hasanah Mohd Ghazali ◽  
Liew Phing Pui
Keyword(s):  
Functional Foods ◽  
Growth Promotion ◽  
Lactobacillus Rhamnosus ◽  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
Preliminary Evaluation ◽  
Lactobacillus Rhamnosus Gg ◽  
Chia Seed ◽  
Seed Mucilage

The characterization of the prebiotic potential of legumes and mucilage are essential and crucial for the development of symbiotic food products. The aim of the present study was to compare and to determine the prebiotic capacity of selected legumes, namely adzuki bean, mung bean, black cowpea, and mucilages from chia seed and flaxseed on the growth of Lactobacillus rhamnosus GG. Resistance towards acid, pancreatin hydrolysis, and the prebiotic scores of the tested substances was determined based on growth promotion after 24 h of incubation. Results revealed that flaxseed mucilage was more resistant against hydrolysis by acid (1.47%) and pancreatin (2.64%) compared to the other samples (5.64 – 18.06% for acid and 10.34 – 15.57% for pancreatin). The relative prebiotic scores for flaxseed mucilage and black cowpea were 98% and 94%, respectively, which were higher than commercial prebiotics including inulin, fructooligosaccharides, and isomaltooligosaccharides. The optimum concentrations of flaxseed mucilage and black cowpea during 36 h of fermentation were 0.8% and 0.4% (w/v), respectively. The findings indicated that flaxseed mucilage was partially digested during gastrointestinal transit and it promotes the growth of the L. rhamnosus GG. The potential prebiotic capacity of flaxseed mucilage and its symbiotic relationship with L. rhamnosus GG suggests that they can be incorporated together for the development of functional foods.

scholarly journals Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherichia coli biofilm formation in co-culture

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Huiyi Song ◽  
Ni Lou ◽  
Jianjun Liu ◽  
Hong Xiang ◽  
Dong Shang
Keyword(s):  
Escherichia Coli ◽  
Proteomic Analysis ◽  
Biofilm Formation ◽  
Lactobacillus Rhamnosus ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Label Free ◽  
Quantitative Proteomic Analysis ◽  
E Coli ◽  
Protein Ubiquitination

Abstract Background Escherichia coli (E. coli) is the principal pathogen that causes biofilm formation. Biofilms are associated with infectious diseases and antibiotic resistance. This study employed proteomic analysis to identify differentially expressed proteins after coculture of E. coli with Lactobacillus rhamnosus GG (LGG) microcapsules. Methods To explore the relevant protein abundance changes after E. coli and LGG coculture, label-free quantitative proteomic analysis and qRT-PCR were applied to E. coli and LGG microcapsule groups before and after coculture, respectively. Results The proteomic analysis characterised a total of 1655 proteins in E. coli K12MG1655 and 1431 proteins in the LGG. After coculture treatment, there were 262 differentially expressed proteins in E. coli and 291 in LGG. Gene ontology analysis showed that the differentially expressed proteins were mainly related to cellular metabolism, the stress response, transcription and the cell membrane. A protein interaction network and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis indicated that the differentiated proteins were mainly involved in the protein ubiquitination pathway and mitochondrial dysfunction. Conclusions These findings indicated that LGG microcapsules may inhibit E. coli biofilm formation by disrupting metabolic processes, particularly in relation to energy metabolism and stimulus responses, both of which are critical for the growth of LGG. Together, these findings increase our understanding of the interactions between bacteria under coculture conditions.

scholarly journals “Masato de Yuca” and “Chicha de Siete Semillas” Two Traditional Vegetable Fermented Beverages from Peru as Source for the Isolation of Potential Probiotic Bacteria

2021 ◽  
Author(s):  
Teresa D. Rebaza-Cardenas ◽  
Kenneth Silva-Cajaleón ◽  
Carlos Sabater ◽  
Susana Delgado ◽  
Nilda D. Montes-Villanueva ◽  
...  
Keyword(s):  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
In Vitro Digestion ◽  
Intestinal Cell ◽  
Adhesion Ability ◽  
Intestinal Cell Line ◽  
Carbohydrate Fermentation ◽  
Traditional Vegetable ◽  
Genetic Profiles

AbstractIn this work, two Peruvian beverages “Masato de Yuca,” typical of the Amazonian communities made from cassava (Manihot esculenta), and “Chicha de Siete Semillas,” made from different cereal, pseudo-cereal, and legume flours, were explored for the isolation of lactic acid bacteria after obtaining the permission of local authorities following Nagoya protocol. From an initial number of 33 isolates, 16 strains with different RAPD- and REP-PCR genetic profiles were obtained. In Chicha, all strains were Lactiplantibacillus plantarum (formerly Lactobacillus plantarum), whereas in Masato, in addition to this species, Limosilactobacillus fermentum (formerly Lactobacillus fermentum), Pediococcus acidilactici, and Weissella confusa were also identified. Correlation analysis carried out with their carbohydrate fermentation patterns and enzymatic profiles allowed a clustering of the lactobacilli separated from the other genera. Finally, the 16 strains were submitted to a static in vitro digestion (INFOGEST model) that simulated the gastrointestinal transit. Besides, their ability to adhere to the human epithelial intestinal cell line HT29 was also determined. Following both procedures, the best probiotic candidate was Lac. plantarum Ch13, a robust strain able to better face the challenging conditions of the gastrointestinal tract and showing higher adhesion ability to the intestinal epithelium in comparison with the commercial probiotic strain 299v. In order to characterize its benefit for human health, this Ch13 strain will be deeply studied in further works.

scholarly journals Autoinducer-2 Triggers the Oxidative Stress Response in Mycobacterium avium, Leading to Biofilm Formation

2008 ◽  
Vol 74 (6) ◽  
pp. 1798-1804 ◽  
Author(s):  
Henriette Geier ◽  
Serge Mostowy ◽  
Gerard A. Cangelosi ◽  
Marcel A. Behr ◽  
Timothy E. Ford
Keyword(s):  
Oxidative Stress ◽  
Stress Response ◽  
Quorum Sensing ◽  
Mycobacterium Avium ◽  
Biofilm Formation ◽  
Transcriptional Response ◽  
Opportunistic Pathogen ◽  
Environmental Stresses ◽  
Oxidative Stress Response ◽  
Autoinducer 2

ABSTRACT Mycobacterium avium is an environmental organism and opportunistic pathogen with inherent resistance to drugs, environmental stresses, and the host immune response. To adapt to these disparate conditions, M. avium must control its transcriptional response to environmental cues. M. avium forms biofilms in various environmental settings, including drinking water pipes and potable water reservoirs. In this study, we investigated the role of the universal signaling molecule autoinducer-2 (AI-2) in biofilm formation by M. avium. The addition of the compound to planktonic M. avium cultures resulted in increased biofilm formation. Microarray and reverse transcriptase PCR studies revealed an upregulation of the oxidative stress response upon addition of AI-2. This suggests that the response to AI-2 might be related to oxidative stress, rather than quorum sensing. Consistent with this model, addition of hydrogen peroxide, a known stimulus of the oxidative stress response, to M. avium cultures resulted in elevated biofilm formation. These results suggest that AI-2 does not act as a quorum-sensing signal in M. avium. Instead, biofilm formation is triggered by environmental stresses of biotic and abiotic origins and AI-2 may exert effects on that level.

scholarly journals YdgG (TqsA) Controls Biofilm Formation in Escherichia coli K-12 through Autoinducer 2 Transport

2006 ◽  
Vol 188 (2) ◽  
pp. 587-598 ◽  
Author(s):  
Moshe Herzberg ◽  
Ian K. Kaye ◽  
Wolfgang Peti ◽  
Thomas K. Wood
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Acid Production ◽  
Polysialic Acid ◽  
Fold Increase ◽  
Uncharacterized Protein ◽  
Biofilm Thickness ◽  
Autoinducer 2 ◽  
Membrane Spanning ◽  
K 12

ABSTRACT YdgG is an uncharacterized protein that is induced in Escherichia coli biofilms. Here it is shown that deletion of ydgG decreased extracellular and increased intracellular concentrations of autoinducer 2 (AI-2); hence, YdgG enhances transport of AI-2. Consistent with this hypothesis, deletion of ydgG resulted in a 7,000-fold increase in biofilm thickness and 574-fold increase in biomass in flow cells. Also consistent with the hypothesis, deletion of ydgG increased cell motility by increasing transcription of flagellar genes (genes induced by AI-2). By expressing ydgG in trans, the wild-type phenotypes for extracellular AI-2 activity, motility, and biofilm formation were restored. YdgG is also predicted to be a membrane-spanning protein that is conserved in many bacteria, and it influences resistance to several antimicrobials, including crystal violet and streptomycin (this phenotype could also be complemented). Deletion of ydgG also caused 31% of the bacterial chromosome to be differentially expressed in biofilms, as expected, since AI-2 controls hundreds of genes. YdgG was found to negatively modulate expression of flagellum- and motility-related genes, as well as other known products essential for biofilm formation, including operons for type 1 fimbriae, autotransporter protein Ag43, curli production, colanic acid production, and production of polysaccharide adhesin. Eighty genes not previously related to biofilm formation were also identified, including those that encode transport proteins (yihN and yihP), polysialic acid production (gutM and gutQ), CP4-57 prophage functions (yfjR and alpA), methionine biosynthesis (metR), biotin and thiamine biosynthesis (bioF and thiDFH), anaerobic metabolism (focB, hyfACDR, ttdA, and fumB), and proteins with unknown function (ybfG, yceO, yjhQ, and yjbE); 10 of these genes were verified through mutation to decrease biofilm formation by 40% or more (yfjR, bioF, yccW, yjbE, yceO, ttdA, fumB, yjiP, gutQ, and yihR). Hence, it appears YdgG controls the transport of the quorum-sensing signal AI-2, and so we suggest the gene name tqsA.

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