Observation of exopolysaccharides (EPS) from Lactobacillus helveticus SBT2171 using the Tokuyasu method

Microscopy ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 286-290
Author(s):  
Takamichi Kamigaki ◽  
Akihiro Ogawa
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Colloidal Gold ◽  
Ruthenium Red ◽  
Lactobacillus Helveticus ◽  
Ruthenium Red Staining ◽  
Observation Method ◽  
Β Lactoglobulin

Abstract Some species of lactic acid bacteria used for the production of natural cheese produce exopolysaccharides (EPS). Electron microscopy is useful for analyzing the microstructure of EPS produced by lactic acid bacteria. However, pretreatments used to observe the microstructure of EPS by electron microscopy, such as dehydration and resin embedding, can result in EPS flowing out easily from the cell. Therefore, in this study, the Tokuyasu method was conducted on cryosection to reduce EPS outflow. Two types of observation method, namely, using lectin and ruthenium red, were conducted in an attempt to observe EPS produced by Lactobacillus helveticus SBT2171. Observation using the lectin method confirmed that colloidal gold particles conjugated with a lectin recognizing β-galactoside were present in the capsule. Structures that appeared to be β-galactoside-containing slime polysaccharides that were released from the cell wall were also observed. Observation using ruthenium red showed that capsular polysaccharides (CPS) in the capsule were present as a net-like structure. Colloidal gold conjugation with an anti-β-lactoglobulin antibody, in addition to ruthenium red staining, allowed the identification of slime polysaccharides released from the cell wall in the milk protein network derived from the culture medium. Based on these results, the Tokuyasu method was considered to be a useful pretreatment method to clarify and observe the presence of EPS. In particular, both CPS in the capsule and slime exopolysaccharides released from the cell wall were visualized.

Antibiotic susceptibility of different lactic acid bacteria strains

2011 ◽  
Vol 2 (4) ◽  
pp. 335-339 ◽  
Author(s):  
N. Karapetkov ◽  
R. Georgieva ◽  
N. Rumyan ◽  
E. Karaivanova
Keyword(s):  
Cell Wall ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Acidophilus ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
Lactobacillus Delbrueckii ◽  
Cell Wall Synthesis ◽  
High Susceptibility ◽  
Lactobacillus Delbrueckii Subsp

Five lactic acid bacteria (LAB) strains belonging to species Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis and Streptococcus thermophilus were tested for their susceptibility to 27 antibiotics. The minimum inhibitory concentrations of each antimicrobial were determined using a microdilution test. Among the strains a high susceptibility was detected for most of the cell-wall synthesis inhibitors (penicillins, cefoxitin and vancomycin) and resistance toward inhibitors of DNA synthesis (trimethoprim/sulfonamides and fluoroquinolones). Generally, the Lactobacillus strains were inhibited by antibiotics such as chloramphenicol, erythromycin and tetracycline at breakpoint levels lower or equal to the levels defined by the European Food Safety Authority. Despite the very similar profile of S. thermophilus LC201 to lactobacilli, the detection of resistance toward erythromycin necessitates the performance of additional tests in order to prove the absence of transferable resistance genes.

The effect of biological additives on the composition and nutritive value of silage

1990 ◽  
Vol 1990 ◽  
pp. 139-139
Author(s):  
M Gonzalez Yanez ◽  
R Mcginn ◽  
D H Anderson ◽  
A R Henderson ◽  
P Phillips
Keyword(s):  
Cell Wall ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lolium Perenne ◽  
Perennial Ryegrass ◽  
Nutritive Value ◽  
Enzyme System ◽  
Lolium Perenne L ◽  
Grass Silage ◽  
Forage Harvester

It Is claimed that the use of the correct enzyme system as an additive on grass silage will satisfactorily control the fermentation and reduce the cell-wall fibre content, thus preserving the nutrients In the silage and aiding their utilisation by the animal (Henderson and McDonald, 1977; Huhtanen et al, 1985; Raurama et al, 1987; Chamberlain and Robertson, 1989; Gordon, 1989;).The aim of the present experiment was to assess the effect of biological additives, enzymes or a combination of enzymes with an Inoculum of lactic acid bacteria, on the composition of silage and on its nutritive value when offered to store lambs as the sole constituent of their diet.On 1st June 1988, first cut perennial ryegrass (Lolium perenne L) at pre-ear emergence was ensiled direct cut untreated (U), treated with a commercial enzyme (E) or with a commercial inoculum of lactic acid bacteria with enzymes (I) in 6t capacity bunker silos. The grass was cut with a mower and lifted with a New Holland precision chop forage harvester. The additives were pumped onto the grass using a dribble bar sited over the pick-up drum.

scholarly journals Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy

ACS Omega ◽  
2021 ◽  
Author(s):  
Johnny Birch ◽  
Sanaullah Khan ◽  
Mikkel Madsen ◽  
Christian Kjeldsen ◽  
Marie Sofie Møller ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Nmr Spectroscopy ◽  
Lactic Acid Bacteria ◽  
Binding Sites ◽  
Β Lactoglobulin

Morphology and ultrastructure of oral strains of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus

1980 ◽  
Vol 30 (2) ◽  
pp. 588-600
Author(s):  
S C Holt ◽  
A C Tanner ◽  
S S Socransky
Keyword(s):  
Electron Microscopy ◽  
Ruthenium Red ◽  
Actinobacillus Actinomycetemcomitans ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Large Numbers ◽  
Haemophilus Aphrophilus ◽  
Ruthenium Red Staining ◽  
Amorphous Surface ◽  
Scanning Electron

Selected human oral and nonoral strains of the genera Actinobacillus and Haemophilus were examined by transmission and scanning electron microscopy. The strains examined were morphologically identical to recognized Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus. By transmission electron microscopy, the cells were typically gram negative in morphology, with several strains possessing some extracellular ruthenium red-staining polymeric material. Numerous vesicular structures, morphologically identical to lipopolysaccharide vesicles, were seen to originate from and be continuous with the surface of the outer membrane. Large numbers of these vesicles were also found in the external environment. Scanning electron microscopic observations revealed that both actinobacilli and haemophili possessed surface projections and an amorphous surface material which connected and covered adjacent cells.

Influence of thermophilic lactic acid bacteria strains on propionibacteria growth and lactate consumption in an Emmental juice-like medium

1999 ◽  
Vol 66 (1) ◽  
pp. 105-113 ◽  
Author(s):  
ANNE THIERRY ◽  
DELPHINE SALVAT-BRUNAUD ◽  
JEAN-LOUIS MAUBOIS
Keyword(s):  
Amino Acids ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Free Amino Acids ◽  
Free Amino ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Helveticus ◽  
The Media ◽  
Lactate Consumption ◽  
Thermophilic Lactic Acid Bacteria

Swiss-type cheeses such as Emmental are characterized by the successive development of thermophilic lactic acid bacteria (TLAB) and propionibacteria. The aim of this study was to determine whether the choice of TLAB strain influenced propionibacteria. TLAB and propionibacteria were cultured sequentially under the conditions prevailing in cheese. Firstly, 11 Emmental juice-like media were prepared by fermenting casein-enriched milk with pure or mixed cultures of TLAB (Lactobacillus helveticus, Lb. delbrueckii subsp. lactis and Streptococcus thermophilus), differing in their proteolytic activities. TLAB cells were then removed by microfiltration. Finally, five strains of Propionibacterium freudenreichii were grown on these media at 24°C under anaerobiosis and their growth characteristics and lactate consumption determined. The media mainly differed in their contents of peptides (1·9–5·3 g/kg) and free amino acids (1·0–5·6 g/kg) and the proportions of lactate isomers (42–92% of the L(+) isomer). Propionibacteria were significantly (P<0·05) influenced by TLAB strains (differences in doubling times of up to 20% and differences in lactate consumption after 600 h culture of up to 52%). The influence of TLAB was similar for all the propionibacteria tested, depended on the TLAB strains and could not be generalized to the TLAB species. Propionibacteria were stimulated by high peptide levels, low levels of free amino acids and NaCl, a low proportion of L(+)-lactate and other undetermined factors. However, variations due to TLAB were less than those between propionibacteria strains.

scholarly journals Design of a Protein-Targeting System for Lactic Acid Bacteria

2001 ◽  
Vol 183 (14) ◽  
pp. 4157-4166 ◽  
Author(s):  
Y. Dieye ◽  
S. Usai ◽  
F. Clier ◽  
A. Gruss ◽  
J.-C. Piard
Keyword(s):  
Cell Wall ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Protein Targeting ◽  
Medium Optimization ◽  
Cell Envelope ◽  
Staphylococcal Nuclease ◽  
Reporter Protein ◽  
Protein Cell ◽  
Export System

ABSTRACT We designed an expression and export system that enabled the targeting of a reporter protein (the staphylococcal nuclease Nuc) to specific locations in Lactococcus lactis cells, i.e., cytoplasm, cell wall, or medium. Optimization of protein secretion and of protein cell wall anchoring was performed with L.lactis cells by modifying the signals located at the N and C termini, respectively, of the reporter protein. Efficient translocation of precursor (∼95%) is obtained using the signal peptide from the lactococcal Usp45 protein and provided that the mature protein is fused to overall anionic amino acids at its N terminus; those residues prevented interactions of Nuc with the cell envelope. Nuc could be covalently anchored to the peptidoglycan by using the cell wall anchor motif of the Streptococcus pyogenes M6 protein. However, the anchoring step proved to not be totally efficient in L. lactis, as considerable amounts of protein remained membrane associated. Our results may suggest that the defect is due to limiting sortase in the cell. The optimized expression and export vectors also allowed secretion and cell wall anchoring of Nuc in food-fermenting and commensal strains of Lactobacillus. In all strains tested, both secreted and cell wall-anchored Nuc was enzymatically active, suggesting proper enzyme folding in the different locations. These results provide the first report of a targeting system in lactic acid bacteria in which the final location of a protein is controlled and biological activity is maintained.

scholarly journals Lyophilized Probiotic Lactic Acid Bacteria Viability in Potato Chips and Its Impact on Oil Oxidation

Foods ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 586
Author(s):  
Heba Mostafa
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fast Food ◽  
Bifidobacterium Longum ◽  
Food Product ◽  
Potato Chips ◽  
Lactobacillus Helveticus ◽  
Live Cells ◽  
Oil Oxidation ◽  
Probiotic Lactic Acid Bacteria

To produce a new probiotic-containing food product, potato chips, as the most preferred fast food, were chosen. Preferably, it should be preserved for a long period without oxidation. The presented study aimed to compare potato chips containing two lyophilized probiotic lactic acid bacteria (Bifidobacterium longum ATCC 15708 and Lactobacillus helveticus LH-B02) in order to retard lipid oxidation. Lyophilization of probiotics was carried out into two cryoprotective media—skim milk (SM) and gelatin/glycerol (GG) as lactose-free medium. Results revealed that GG and SM media were the most suitable for lyophilization of B. longum and L. helveticus, respectively. The lyophilized live cells were incorporated in potato chips, packed and their effect on oil oxidation was assessed. Results showed that the lyophilized B. longum in SM remained alive at 6.5 log CFU/g for 4 months at 30 °C. Interestingly, potato chip bags containing B. longum lyophilized in SM medium exhibited a decrease in peroxide value (PV) and acid value (AV) of the extracted oil by 40.13% and 25%, respectively, compared to the control bags. The created probiotic potato chips containing B. longum fulfill the criteria of the probiotic product besides the prime quality and sensory attributes.

Sign in / Sign up

Export Citation Format

Share Document