scholarly journals Bioluminescence Imaging Study of Spatial and Temporal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice

2012 ◽  
Vol 79 (4) ◽  
pp. 1086-1094 ◽  
Author(s):  
Catherine Daniel ◽  
Sabine Poiret ◽  
Véronique Dennin ◽  
Denise Boutillier ◽  
Bruno Pot
Keyword(s):  
Lactic Acid ◽  
Gastrointestinal Tract ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactococcus Lactis ◽  
Bioluminescence Imaging ◽  
Ex Vivo ◽  
Mammalian Host ◽  
Content Type

ABSTRACTLactic acid bacteria, especially lactobacilli, are common inhabitants of the gastrointestinal tract of mammals, for which they have received considerable attention due to their putative health-promoting properties. In this study, we describe the development and application of luciferase-expressingLactobacillus plantarumandLactococcus lactisstrains for noninvasivein vivomonitoring in the digestive tract of mice. We report for the first time the functionalin vitroexpression inLactobacillus plantarumNCIMB8826 and inLactococcus lactisMG1363 of the click beetle luciferase (CBluc), as well asGaussiaand bacterial luciferases, using a combination of vectors, promoters, and codon-optimized genes. We demonstrate that a CBluc construction is the best-performing luciferase system for the noninvasivein vivodetection of lactic acid bacteria after oral administration. The persistence and viability of both strains was studied by bioluminescence imaging in anesthetized mice and in mouse feces.In vivobioluminescence imaging confirmed that after a single or multiple oral administrations,L. lactishas shorter survival times in the mouse gastrointestinal tract thanL. plantarum, and it also revealed the precise gut compartments where both strains persisted. The application of luciferase-labeled bacteria has significant potential to allow thein vivoandex vivostudy of the interactions of lactic acid bacteria with their mammalian host.

scholarly journals Dual-Color Bioluminescence Imaging for Simultaneous Monitoring of the Intestinal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice

2015 ◽  
Vol 81 (16) ◽  
pp. 5344-5349 ◽  
Author(s):  
Catherine Daniel ◽  
Sabine Poiret ◽  
Véronique Dennin ◽  
Denise Boutillier ◽  
Delphine Armelle Lacorre ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Gastrointestinal Tract ◽  
Lactic Acid Bacteria ◽  
Bioluminescence Imaging ◽  
Ex Vivo ◽  
Spectral Unmixing ◽  
Content Type ◽  
Dual Color ◽  
Click Beetle

ABSTRACTLactic acid bacteria are found in the gastrointestinal tract of mammals and have received tremendous attention due to their health-promoting properties. We report the development of two dual-color luciferase-producingLactobacillus(Lb.)plantarumandLactococcus(Lc.)lactisstrains for noninvasive simultaneous tracking in the mouse gastrointestinal tract. We previously described the functional expression of the red luciferase mutant (CBRluc) fromPyrophorus plagiophthalamusinLb. plantarumNCIMB8826 andLc. lactisMG1363 (C. Daniel, S. Poiret, V. Dennin, D. Boutillier, and B. Pot, Appl Environ Microbiol79:1086–1094, 2013,http://dx.doi.org/10.1128/AEM.03221-12). In this study, we determined that CBRluc is a better-performing luciferase forin vivolocalization of both lactic acid bacteria after oral administration than the green click beetle luciferase mutant construct developed in this study. We further established the possibility to simultaneously detect red- and green-emitting lactic acid bacteria by dual-wavelength bioluminescence imaging in combination with spectral unmixing. The difference in spectra of light emission by the red and green click beetle luciferase mutants and dual bioluminescence detection allowedin vitroandin vivoquantification of the red and green emitted signals; thus, it allowed us to monitor the dynamics and fate of the two bacterial populations simultaneously. Persistence and viability of both strains simultaneously administered to mice in different ratios was studiedin vivoin anesthetized mice andex vivoin mouse feces. The application of dual-luciferase-labeled bacteria has considerable potential to simultaneously study the interactions and potential competitions of different targeted bacteria and their hosts.

scholarly journals Evaluation of the Efficiency of Ethanol Precipitation and Ultrafiltration on the Purification and Characteristics of Exopolysaccharides Produced by Three Lactic Acid Bacteria

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Manel Ziadi ◽  
Taroub Bouzaiene ◽  
Sana M’Hir ◽  
Kaouther Zaafouri ◽  
Ferid Mokhtar ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactococcus Lactis ◽  
Leuconostoc Mesenteroides ◽  
Ftir Analysis ◽  
Molecular Weights ◽  
Molar Ratios ◽  
Monomer Composition ◽  
Ethanol Precipitation

Exopolysaccharides (EPS) produced by three Lactic Acid Bacteria strains,Lactococcus lactisSLT10,Lactobacillus plantarumC7, andLeuconostoc mesenteroidesB3, were isolated using two methods: ethanol precipitation (EPS-ETOH) and ultrafiltration (EPS-UF) through a 10 KDa cut-off membrane. EPS recovery by ultrafiltration was higher than ethanol precipitation forLactococcus lactisSLT10 andLactobacillus plantarumC7. However, it was similar with both methods forLeuconostoc mesenteroidesB3. The monomer composition of the EPS fractions revealed differences in structures and molar ratios between the two studied methods. EPS isolated fromLactococcus lactisSLT10 are composed of glucose and mannose for EPS-ETOH against glucose, mannose, and rhamnose for EPS-UF. EPS extracted fromLactobacillus plantarumC7 andLeuconostoc mesenteroidesB3 showed similar composition (glucose and mannose) but different molar ratios. The molecular weights of the different EPS fractions ranged from 11.6±1.83 to 62.4±2.94 kDa. Molecular weights of EPS-ETOH fractions were higher than those of EPS-UF fractions. Fourier transform infrared (FTIR) analysis revealed a similarity in the distribution of the functional groups (O-H, C-H, C=O, -COO, and C-O-C) between the EPS isolated from the three strains.

scholarly journals Application of some lactic acid bacteria strains to improve fermentation and aerobic stability of maize silage

2019 ◽  
Vol 7 (1-2) ◽  
pp. 127-132
Author(s):  
Judit Peter Szucs ◽  
Agnes Suli ◽  
Timea Suli Zakar ◽  
Elizabet Berecz ◽  
Mate Pek
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactococcus Lactis ◽  
Enterococcus Faecium ◽  
Acid Production ◽  
Untreated Control ◽  
Raw Material ◽  
Aerobic Stability ◽  
Whole Plant

The object of the trial was to study the effect of some lactic acid bacteria strains on the fermentation and aerobic stability of whole plant maize silages.The whole plant maize raw material was 32% DM, in soft cheddar stage of grain ripeness. It was ensiled in 4.2 litre capacity glass micro-size silos in 5 replicates /each treatment and stored on constant air conditioned room temperature (22 oC) during 95 days. The average packing density of raw material was 211 kg DM/m3.The applied treatments: 1. Untreated control maize, 2. Enterococcus faecium 100,000 CFU/g fresh maize (FM), 3. Lactobacillus plantarum 50,000 CFU/g FM + Enterococcus faecium 50,000 CFU/g FM, 4. Lactococcus lactis 100,000 CFU/g FM, 5. Lactobacillus plantarum 50,000 CFU/g FM + Lactococcus lactis 50,000 CFU/g FM, 6. Lactobacillus plantarum 100,000 CFU/g FM.Aerobic stability study:  Applied Honig (1990 system).The main experiences are the following: Applied lactic acid bacteria strains improved the quality of fermentation of maize in general compare to untreated control one.Lactic acid bacteria strains significantly stimulated lactic acid production and decreased propionic and butyric acid production. The origin of ammonia decreased also under influence of lactic acid bacteria strains in unaerobic conditions.Enterococcus faecium and.Lactococcus lactis are not able to protect the maize silages against the aerobic deterioration with the applied dosage.  Lactobacillus plantarum itself produced the most favourable fermentation characteristics and protected the aerobic stability of silage the most effectively (during 4 day) compare to all other treated maize silages.

scholarly journals Versatile Cas9-Driven Subpopulation Selection Toolbox forLactococcus lactis

2018 ◽  
Vol 84 (8) ◽  
Author(s):  
Simon van der Els ◽  
Jennelle K. James ◽  
Michiel Kleerebezem ◽  
Peter A. Bron
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactococcus Lactis ◽  
Mobile Genetic Elements ◽  
Broad Host Range ◽  
Content Type ◽  
Genetic Elements ◽  
Wide Range ◽  
Dairy Fermentation ◽  
Selection Of

ABSTRACTCRISPR-Cas9 technology has been exploited for the removal or replacement of genetic elements in a wide range of prokaryotes and eukaryotes. Here, we describe the extension of the Cas9 application toolbox to the industrially important dairy speciesLactococcus lactis. The Cas9 expression vector pLABTarget, encoding theStreptocccus pyogenesCas9 under the control of a constitutive promoter, was constructed, allowing plug and play introduction of short guide RNA (sgRNA) sequences to target specific genetic loci. Introduction of apepN-targeting derivative of pLABTarget intoL. lactisstrain MG1363 led to a strong reduction in the number of transformants obtained, which did not occur in apepNdeletion derivative of the same strain, demonstrating the specificity and lethality of the Cas9-mediated double-strand breaks in the lactococcal chromosome. Moreover, the same pLABTarget derivative allowed the selection of apepNdeletion subpopulation from its corresponding single-crossover plasmid integrant precursor, accelerating the construction and selection of gene-specific deletion derivatives inL. lactis. Finally, pLABTarget, which contained sgRNAs designed to target mobile genetic elements, allowed the effective curing of plasmids, prophages, and integrative conjugative elements (ICEs). These results establish that pLABTarget enables the effective exploitation of Cas9 targeting inL. lactis, while the broad-host-range vector used suggests that this toolbox could readily be expanded to other Gram-positive bacteria.IMPORTANCEMobile genetic elements inLactococcus lactisand other lactic acid bacteria (LAB) play an important role in dairy fermentation, having both positive and detrimental effects during the production of fermented dairy products. The pLABTarget vector offers an efficient cloning platform for Cas9 application in lactic acid bacteria. Targeting Cas9 toward mobile genetic elements enabled their effective curing, which is of particular interest in the context of potentially problematic prophages present in a strain. Moreover, Cas9 targeting of other mobile genetic elements enables the deciphering of their contribution to dairy fermentation processes and further establishment of their importance for product characteristics.

Differential Sensitivity of Clostridium botulinum Strains to Nisin is Not Biotype-Associated

1992 ◽  
Vol 55 (6) ◽  
pp. 444-448 ◽  
Author(s):  
THOMAS J. MONTVILLE ◽  
ANN M. ROGERS ◽  
AMECHI OKEREKE
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactococcus Lactis ◽  
Yeast Extract ◽  
Clostridium Botulinum ◽  
Differential Sensitivity ◽  
Vegetative Cells ◽  
Significant Difference ◽  
Peptone Yeast Extract Glucose

The sensitivities of proteolytic and nonproteolytic Clostridium botulinum strains to nisin and other bacteriocins were investigated. Although there were statistically different nisin sensitivities among vegetative cells from 18 C. botulinum strains, these differences were not biotype-associated. When inoculated into tryptose peptone yeast extract glucose broth containing nisin at various levels, spores from strain 56 A were not inhibited at all by 100 IU/ml of nisin. About 2,500 IU/ml was required to inhibit growth for 30 d. In contrast, only 10 IU/ml was required to inhibit strain 169 for 30 d. Both strains were completely inhibited at the 10,000 IU/ml limit allowed in processed cheeses. Spores from strains having limited (strain 56A), moderate (strain 25675), or extreme (strain 169) nisin sensitivity were examined for bacteriocin-mediated inhibition by Lactococcus lactis 11454, Pediococcus pentosaceus 43200, P. pentosaceus 43201, and Lactobacillus plantarum BN using the spot-on-the-lawn method. While the differences in nisin sensitivity were confirmed, there was no statistically significant difference in their sensitivities to the bacteriocins produced by other lactic acid bacteria.

scholarly journals An Extracellular Serine/Threonine-Rich Protein from Lactobacillus plantarum NCIMB 8826 Is a Novel Aggregation-Promoting Factor with Affinity to Mucin

2013 ◽  
Vol 79 (19) ◽  
pp. 6059-6066 ◽  
Author(s):  
Arancha Hevia ◽  
Noelia Martínez ◽  
Víctor Ladero ◽  
Miguel A. Álvarez ◽  
Abelardo Margolles ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
D1 Protein ◽  
Extracellular Proteins ◽  
Human Immune System ◽  
Content Type ◽  
Probiotic Lactic Acid Bacteria ◽  
Adhesion Role ◽  
Internal Fragment

ABSTRACTAutoaggregation in lactic acid bacteria is directly related to the production of certain extracellular proteins, notably, aggregation-promoting factors (APFs). Production of aggregation-promoting factors confers beneficial traits to probiotic-producing strains, contributing to their fitness for the intestinal environment. Furthermore, coaggregation with pathogens has been proposed to be a beneficial mechanism in probiotic lactic acid bacteria. This mechanism would limit attachment of the pathogen to the gut mucosa, favoring its removal by the human immune system. In the present paper, we have characterized a novel aggregation-promoting factor inLactobacillus plantarum. A mutant with a knockout of the D1 gene showed loss of its autoaggregative phenotype and a decreased ability to bind to mucin, indicating an adhesion role of this protein. In addition, heterologous production of the D1 protein or an internal fragment of the protein, characterized by its abundance in serine/threonine, strongly induced autoaggregation inLactococcus lactis. This result strongly suggested that this internal fragment is responsible for the bioactivity of D1 as an APF. To our knowledge, this is the first report on a gene coding for an aggregation-promoting factor inLb. plantarum.

scholarly journals CRISPR/Cas9-Assisted Seamless Genome Editing in Lactobacillus plantarum and Its Application in N-Acetylglucosamine Production

2019 ◽  
Vol 85 (21) ◽  
Author(s):  
Ding Zhou ◽  
Zhennan Jiang ◽  
Qingxiao Pang ◽  
Yuan Zhu ◽  
Qian Wang ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Genome Editing ◽  
Lactobacillus Plantarum ◽  
Strain Engineering ◽  
Single Stranded Dna ◽  
Content Type ◽  
Recombination Efficiency ◽  
Double Stranded Dna ◽  
Exogenous Genes

ABSTRACT Lactobacillus plantarum is a potential starter and health-promoting probiotic bacterium. Effective, precise, and diverse genome editing of Lactobacillus plantarum without introducing exogenous genes or plasmids is of great importance. In this study, CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering was established in L. plantarum WCFS1 to seamlessly edit the genome, including gene knockouts, insertions, and point mutations. To optimize our editing method, phosphorothioate modification was used to improve the dsDNA insertion, and adenine-specific methyltransferase was used to improve the ssDNA recombination efficiency. These strategies were applied to engineer L. plantarum WCFS1 toward producing N-acetylglucosamine (GlcNAc). nagB was truncated to eliminate the reverse reaction of fructose-6-phosphate (F6P) to glucosamine 6-phosphate (GlcN-6P). Riboswitch replacement and point mutation in glmS1 were introduced to relieve feedback repression. The resulting strain produced 797.3 mg/liter GlcNAc without introducing exogenous genes or plasmids. This strategy may contribute to the available methods for precise and diverse genetic engineering in lactic acid bacteria and boost strain engineering for more applications. IMPORTANCE CRISPR/Cas9-assisted recombineering is restricted in lactic acid bacteria because of the lack of available antibiotics and vectors. In this study, a seamless genome editing method was carried out in Lactobacillus plantarum using CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering, and recombination efficiency was effectively improved by endogenous adenine-specific methyltransferase overexpression. L. plantarum WCFS1 produced 797.3 mg/liter N-acetylglucosamine (GlcNAc) through reinforcement of the GlcNAc pathway, without introducing exogenous genes or plasmids. This seamless editing strategy, combined with the potential exogenous GlcNAc-producing pathway, makes this strain an attractive candidate for industrial use in the future.

scholarly journals Enantioselective Regulation of Lactate Racemization by LarR in Lactobacillus plantarum

2014 ◽  
Vol 197 (1) ◽  
pp. 219-230 ◽  
Author(s):  
Benoît Desguin ◽  
Philippe Goffin ◽  
Nordine Bakouche ◽  
Aurélie Diman ◽  
Eric Viaene ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactobacillus Plantarum ◽  
Point Mutations ◽  
Racemic Mixture ◽  
Content Type ◽  
Sugar Fermentation ◽  
Gel Retardation ◽  
Positive Effect

Lactobacillus plantarumis a lactic acid bacterium that produces a racemic mixture ofl- andd-lactate from sugar fermentation. The interconversion of lactate isomers is performed by a lactate racemase (Lar) that is transcriptionally controlled by thel-/d-lactate ratio and maximally induced in the presence ofl-lactate. We previously reported that the Lar activity depends on the expression of two divergently oriented operons: (i) thelarABCDEoperon encodes the nickel-dependent lactate racemase (LarA), its maturases (LarBCE), and a lactic acid channel (LarD), and (ii) thelarR(MN)QOoperon encodes a transcriptional regulator (LarR) and a four-component ABC-type nickel transporter [Lar(MN), in which the M and N components are fused, LarQ, and LarO]. LarR is a novel regulator of the Crp-Fnr family (PrfA group). Here, the role of LarR was further characterizedin vivoandin vitro. We show that LarR is a positive regulator that is absolutely required for the expression of Lar activity. Using gel retardation experiments, we demonstrate that LarR binds to a 16-bp palindromic sequence (Lar box motif) that is present in thelarR-larAintergenic region. Mutations in the Lar box strongly affect LarR binding and completely abolish transcription from thelarApromoter (PlarA). Two half-Lar boxes located between the Lar box and the −35 box of PlarApromote LarR multimerization on DNA, and point mutations within one or both half-Lar boxes inhibit PlarAinduction byl-lactate. Gel retardation and footprinting experiments indicate thatl-lactate has a positive effect on the binding and multimerization of LarR, whiled-lactate antagonizes the positive effect ofl-lactate. A possible mechanism of LarR regulation by lactate enantiomers is proposed.

Antigenotoxic properties of lactic acid bacteria in vivo in the gastrointestinal tract of rats

1993 ◽  
Vol 20 (3) ◽  
pp. 271-281 ◽  
Author(s):  
Beatrice L. Pool‐Zobel ◽  
Barbara Bertram ◽  
Martin Knoll ◽  
Renate Lambertz ◽  
Christian Neudecker ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Gastrointestinal Tract ◽  
Lactic Acid Bacteria

scholarly journals Development of the Traditional Tape Ketan Into Probiotic Drink

2018 ◽  
Vol 15 (1) ◽  
pp. 11
Author(s):  
Endang S. Rahayu ◽  
Asri Nursiwi ◽  
Bedri Sekar N ◽  
Supriyanto Supriyanto
Keyword(s):  
Lactic Acid ◽  
Gastrointestinal Tract ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Health Benefit ◽  
Viable Count ◽  
Inlet Temperature ◽  
Spray Dryer ◽  
Glutinous Rice ◽  
Probiotic Lactobacillus

Probiotics are living microorganism which give health benefit when consumed. Probiotic needs a ‘vehicle’ for their specialized actions in gastrointestinal tract. In this research, tape ketan was used as ‘vehicle’. The objective of this research was to produce probiotic tape ketan powder as an ingredient of functional beverage. Tape ketan used in this research was made by fermentation of glutinous rice with ragi and supplemented with Lactobacillus plantarum Dad 13 (107CFU/g glutinous rice). Probiotic powder was obtained using spray dryer with inlet temperature of 90oC with the addition of 35% maltodextrin. The powder was reconstituted into water with addition of pectin and sucrose. The results showed that viability of lactic acid bacteria and L. plantarum decreased after drying (0.82 and 0.90 log cycle respectively) with viable count from 1.29x1010 CFU to 1.96x109 CFU for lactic acid bacteria and from 1.04x1010 CFU to 1.32x109 CFU for L. plantarum. To obtain probiotic tape ketan beverage, 20% tape ketan powder (w/v), 0.5% pectin (w/v) and 4% sucrose (w/v) were reconstituted. Tape ketan powder which was supplemented with probiotic Lactobacillus plantarum Dad 13 is potential as an ingredient of functional beverage (from the viability after drying).

Sign in / Sign up

Export Citation Format

Share Document