scholarly journals Mucosal Adhesion Properties of the Probiotic Lactobacillus rhamnosus GG SpaCBA and SpaFED Pilin Subunits

2010 ◽  
Vol 76 (7) ◽  
pp. 2049-2057 ◽  
Author(s):  
Ingemar von Ossowski ◽  
Justus Reunanen ◽  
Reetta Satokari ◽  
Satu Vesterlund ◽  
Matti Kankainen ◽  
...  

ABSTRACT Lactobacillus rhamnosus GG is a well-established Gram-positive probiotic strain, whose health-benefiting properties are dependent in part on prolonged residence in the gastrointestinal tract and are likely dictated by adherence to the intestinal mucosa. Previously, we identified two pilus gene clusters (spaCBA and spaFED) in the genome of this probiotic bacterium, each of which contained the predicted genes for three pilin subunits and a single sortase. We also confirmed the presence of SpaCBA pili on the cell surface and attributed an intestinal mucus-binding capacity to one of the pilin subunits (SpaC). Here, we report cloning of the remaining pilin genes (spaA, spaB, spaD, spaE, and spaF) in Escherichia coli, production and purification of the recombinant proteins, and assessment of the adherence of these proteins to human intestinal mucus. Our findings indicate that the SpaB and SpaF pilin subunits also exhibit substantial binding to mucus, which can be inhibited competitively in a dose-related manner. Moreover, the binding between the SpaB pilin subunit and the mucosal substrate appears to operate through electrostatic contacts and is not related to a recognized mucus-binding domain. We conclude from these results that it is conceivable that two pilin subunits (SpaB and SpaC) in the SpaCBA pilus fiber play a role in binding to intestinal mucus, but for the uncharacterized and putative SpaFED pilus fiber only a single pilin subunit (SpaF) is potentially responsible for adhesion to mucus.

2010 ◽  
Vol 105 (6) ◽  
pp. 887-894 ◽  
Author(s):  
Łukasz Grześkowiak ◽  
Erika Isolauri ◽  
Seppo Salminen ◽  
Miguel Gueimonde

Production and manufacturing methods and the food carrier may influence the properties of probiotic strains, and have an impact on the outcome of clinical intervention studies. The aim of the present study was to establish whether the properties of a specific probiotic strain,Lactobacillus rhamnosusGG, may differ depending on the product and source of the strain. In total, fifteen differentL. rhamnosusisolates, among them fourteen labelled asL.rhamnosusGG, were isolated from specific probiotic products. The micro-organisms were phenotypically and genotypically characterised. Their adhesion properties were compared using the human intestinal mucus model, and the ability of the isolates to influence model pathogen adhesion to human colonic mucus was assessed. AllL. rhamnosusisolates used were confirmed as members of the speciesL. rhamnosus. Except the reference strain OL, allL.rhamnosusisolates showed randomly amplified polymorphic DNA, enterobacterial repetitive intergenic consensus and pulsed-field gel electrophoresis profiles identical to that ofL. rhamnosusGG (ATCC 53103). AllL.rhamnosusisolates showed similar tolerance to acid and were able to bind to human colonic mucus. However, pathogen exclusion by inhibition and competition varied significantly among the differentL. rhamnosusisolates and pathogens tested. The results suggest that different sources of the same probiotic may have significantly altered strain properties. This should be considered inin vivostudies on human subjects and also for quality control of probiotic products.


2009 ◽  
Vol 123 (2) ◽  
pp. S200-S200
Author(s):  
R.J. Boyle ◽  
L. Mah ◽  
S. Kivivuori ◽  
A. Chen ◽  
S.J. Lahtinen ◽  
...  

2013 ◽  
Vol 67 (7) ◽  
Author(s):  
Ľubomír Valík ◽  
Alžbeta Medveďová ◽  
Michal Čižniar ◽  
Denisa Liptáková

AbstractThe application of secondary temperature models on growth rates of Lactobacillus rhamnosus GG, the much studied probiotic bacterium, is investigated. Growth parameters resulting from a primary fitting were modelled against temperature using the following models: Hinshelwood model (H), Ratkowsky extended model (RTK2), Zwietering model (ZWT), and cardinal temperature model with inflection (CTMI). As experienced by other authors, the RTK2, ZWT, and CTMI models provided the best statistical indices related to fitting the experimental data. Moreover, with the biological background, the following cardinal temperatures of L. rhamnosus GG resulted from the study by the model application: t min = 2.7°C, t opt = 44.4°C, t max = 52.0°C. The growth rate of the strain under study at optimal temperature was 0.88 log10(CFU mL−1 h−1).


Foods ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1252
Author(s):  
Kandasamy Saravanakumar ◽  
Anbazhagan Sathiyaseelan ◽  
Arokia Vijaya Anand Mariadoss ◽  
Ramachandran Chelliah ◽  
Xiaowen Hu ◽  
...  

This work analyzed the individual and combined effects of biochemical additives and probiotic strain Lactobacillus rhamnosus GG on red and yellow fresh-cut bell pepper (R- and Y-FCBP, respectively) stored at two different temperatures (4 °C and 15 °C) for 15 days. The results revealed that the combined application of biochemical additives and L. rhamnosus GG inhibited the colonization of total bacterial counts (25.10%), total Salmonella counts (38.32%), total Listeria counts (23.75%), and total fungal counts (61.90%) in FCBP. Total bacterial colonization was found to be higher in R-FCBP (1188.09 ± 9.25 CFU g−1) than Y-FCBP (863.96 ± 7.21 CFU g−1). The storage at 4 °C was prevented 35.38% of microbial colonization in FCBP. Importantly, the L. rhamnosus GG count remained for up to 12 days. Moreover, the combined inoculation of the biochemical additives and L. rhamnosus GG treatments (T3) maintained the quality of R- and Y-FCBP for up to 12 days at 4 °C without any loss of antioxidant properties. This work reports the successful utilization of L. rhamnosus GG as a preservative agent for maintaining the quality of FCBP by preventing microbial colonization.


2006 ◽  
Vol 69 (8) ◽  
pp. 2011-2015 ◽  
Author(s):  
PATRICIA RUAS-MADIEDO ◽  
MIGUEL GUEIMONDE ◽  
ABELARDO MARGOLLES ◽  
CLARA G. de los REYES-GAVILÁN ◽  
SEPPO SALMINEN

Exopolysaccharides (EPSs) are exocellular polymers present in the surface of many bacteria, including Lactobacillus and Bifidobacterium. The genome sequence of several strains revealed the presence of EPS-encoding genes. However, the physiological role that EPSs play in the bacterial ecology still remains uncertain. In this study, we have assessed the effect of EPSs produced by Lactobacillus rhamnosus GG, Bifidobacterium longum NB667, and Bifidobacterium animalis IPLA-R1 on the adhesion of probiotic and enteropathogen strains to human intestinal mucus. The EPS fraction GG had no significant effect on the adhesion of L. rhamnosus GG and B. animalis IPLA-R1. However, the EPS fractions NB667 and IPLA-R1 significantly reduced the adherence of both probiotic strains. In contrast, the three EPS fractions increased the adhesion of Enterobacter sakazakii ATCC 29544 and Escherichia coli NCTC 8603. Higher adherence of Salmonella enterica serovar Typhimurium ATCC 29631 and Clostridium difficile ATCC 9689 was detected in the presence of the EPS fractions GG and NB667. In general, these effects were obtained at EPS concentrations of up to 5 mg/ml, and they were EPS dose dependent. The competitive exclusion of probiotics in the presence of EPS could suggest the involvement of these biopolymers in the adhesion to mucus. The increase in the adherence of enteropathogens could be explained if components of the pathogen surface are able to bind to specific EPSs and the bound EPSs are able to adhere to mucus. To the best of our knowledge, this is the first work reporting the effect of EPSs from probiotics on bacterial adhesion properties.


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

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.


2020 ◽  
Vol 103 (6) ◽  
pp. 1604-1609 ◽  
Author(s):  
Hanan R Shehata ◽  
Steven G Newmaster

Abstract Background Strain Lactobacillus rhamnosus GG is one of the best-studied and most widely used probiotic strains, with various health benefits. Because probiotic health benefits and safety are strain specific, the availability of a reliable assay for specific identification of Lactobacillus rhamnosus GG is vital to ensure probiotic efficacy. Objective To design and validate a probe-based real-time PCR assay for specific identification of strain Lactobacillus rhamnosus GG. Method Rapid Annotation using Subsystem Technology (RAST) was used to find a unique sequence region in the genome of Lactobacillus rhamnosus GG. A probe-based assay was designed and evaluated for specificity, sensitivity, efficiency, repeatability, and reproducibility. Results RAST identified a unique gene coding for a hypothetical protein in the genome of Lactobacillus rhamnosus GG. The assay successfully amplified all 22 target samples and did not amplify any of the 28 non-target strains, achieving 100% true positive and 0% false positive results. The Limit of Detection (LOD) was determined to be 0.001 ng. Reaction efficiency values, from three dilution series, were 96.4%, 93.3%, and 96.8% with R square values of 0.9974, 0.9981, and 0.9998, respectively. Relative standard deviation (RSD, %) of repeatability was below 1% and RSD of reproducibility was below 4%. Conclusions This Lactobacillus rhamnosus GG specific assay proved to be specific, sensitive, efficient, and reproducible. Since the assay was evaluated on two real-time PCR platforms, including a portable one, the assay can be used for onsite testing throughout the supply chain. Highlights The availability of validated and reliable assays for strain-specific identification plays a vital role in achieving compliance in probiotic products.


Author(s):  
Michał Świeca ◽  
Monika Kordowska-Wiater ◽  
Monika Pytka ◽  
Łukasz Sęczyk ◽  
Urszula Gawlik-Dziki

Probiotics and prebiotics play an important role in human and animal nutrition. Those research studies were performed to evaluate the potential of using legume sprouts as carriers for probiotic strain of Lactobacillus rhamnosus GG. They determined the effect of legume species, temperature of sprouting, and inoculation methods of seeds or growing sprouts on the survival and/or growth of probiotics. It was found that the count of bacteria in sprouts depended on the germination temperature, inoculation methods as well as on the species of legume used as a carrier. The beans examined (Adzuki and Mung) germinated effectively at a temperature between 25 ÷ 35 ºC. And the lentil sprouted most effectively at 25 ºC. In the case of soy-bean and lentil, the temperature of 35 ºC caused the germination efficiency to decrease. The growth of Lb. rhamnosus GG was reported only in the case of the lentil and soy-bean sprouts obtained from the seeds imbibed in an inoculum and germinated at 25 ºC. The count of probiotic bacteria was 3.1×106 and 7.18×106 CFU per grams of fresh mass, respectively. The sprouts obtained from the bean seeds analyzed did not provide any conditions for probiotic bacteria to survive and grow. The best carrier for the probiotic bacteria studied were the soy-bean sprouts; in their case, after inoculation of seeds and using a suspension of probiotic bacteria, the sprouts obtained at 25 ºC had the best quality parameters.


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