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The effect of cold plasma (CP) on phenolic compound (PC) and biogenic amine (BA) contents of red wine was investigated for the first time. The influence of CP was compared with the effects of a wine preservation using potassium metabisulfite and a combined method. The PC profile was determined by UPLC-PDA-MS/MS while BAs using DLLME-GC-MS. Chemometric analysis also was used. The content of PCs was 3.1% higher in the sample preserved by CP treatment (5 min, helium/nitrogen) compared to a sample preserved by the addition of potassium metabisulfite (100 mg/L). On a positive note, CP treatment reduced the concentration of BAs in the wine samples. The lowest BA contents were recorded after 10 min of cold plasma (helium/oxygen) treatment with the addition of potassium metabisulfite (1120.85 μg/L). The results may promote interest in CP as a potential alternative method for the preservation of wine and other alcoholic beverages.

Fireweed (Epilobium angustifolium L.) has attracted attention due to its numerous biological activities. In the present work, we produced a polyphenol-rich freeze-dried infusion (407.02 ± 7.10 mg GAE/g) in order to measure, for the first time, the evolution of anticholinesterase activity during in vitro digestion. During the ‘gastric’ phase, there was an increase in the inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity followed by a decrease during the ‘small intestinal’ phase. A strong correlation between total polyphenolic content (TPC) and the anticholinesterase activity of the digested fluids was observed (r = 0.90 for TPC/AChE; r = 0.88 for TPC/BChE, p

Scharakteryzowano właściwości, strukturę oraz spektrum aktywności plantarycyn wytwarzanych przez niektóre szczepy Lactobacillus. Przedstawiono ich klasyfikację, mechanizm biosyntezy, modele działania na inne bakterie oraz stabilność. Plantarycyny są dość różnorodne pod względem właściwości fizykochemicznych, struktury i spektrum aktywności przeciwdrobnoustrojowej. Szczepy Lactobacillus plantarum zdolne do syntezy plantarycyn należą do mikroorganizmów najczęściej obecnych w fermentowanych produktach spożywczych, a także w napojach i innych wyrobach z dodatkiem mleka. Budowa cząsteczek, ich stabilność oraz mechanizm działania sprawiają, że plantarycyny charakteryzują się skutecznym działaniem bakteriobójczym. Ze względu na niewielką oporność organizmów patogennych na te substancje mogą być one alternatywą dla wielu stosowanych obecnie związków o działaniu przeciwdrobnoustrojowym. Plantarycyny wpływają na zahamowanie wzrostu drobnoustrojów chorobotwórczych, a w konsekwencji na poprawę zdrowia człowieka. Z uwagi na coraz większe zainteresowanie produktami naturalnymi plantarycyny mogą w przyszłości znaleźć zastosowanie w różnych gałęziach przemysłu. Wyniki badań dotyczące ich wykorzystania mogą stać się podstawą do projektowania efektywnych preparatów probiotycznych, kultur starterowych do produkcji fermentowanej żywności czy nowych metod zabezpieczania żywności, co w konsekwencji wpłynie na poprawę zdrowia i jakości życia człowieka.

Microorganisms have been harnessed to process raw plants into fermented foods. The adaptation to a variety of plant environments has resulted in a nearly inseparable association between the bacterial species and the plant with a characteristic chemical profile. Lactic acid bacteria, which are known for their ability to adapt to nutrient-rich niches, have altered their genomes to dominate specific habitats through gene loss or gain. Molecular biology approaches provide a deep insight into the evolutionary process in many bacteria and their adaptation to colonize the plant matrix. Knowledge of the adaptive characteristics of microorganisms facilitates an efficient use thereof in fermentation to achieve desired final product properties. With their ability to acidify the environment and degrade plant compounds enzymatically, bacteria can modify the textural and organoleptic properties of the product and increase the bioavailability of plant matrix components. This article describes selected microorganisms and their competitive survival and adaptation in fermented fruit and vegetable environments. Beneficial changes in the plant matrix caused by microbial activity and their beneficial potential for human health are discussed as well.

The production of mead holds great value for the Polish liquor industry, which is why the bacterium that spoils mead has become an object of concern and scientific interest. This article describes, for the first time, Lactobacillus hilgardii FLUB newly isolated from mead, as a mead spoilage bacteria. Whole genome sequencing of L. hilgardii FLUB revealed a 3 Mbp chromosome and five plasmids, which is the largest reported genome of this species. An extensive phylogenetic analysis and digital DNA-DNA hybridization confirmed the membership of the strain in the L. hilgardii species. The genome of L. hilgardii FLUB encodes 3043 genes, 2871 of which are protein coding sequences, 79 code for RNA, and 93 are pseudogenes. L. hilgardii FLUB possesses three clustered regularly interspaced short palindromic repeats (CRISPR), eight genomic islands (44,155 bp to 6345 bp), and three (two intact and one incomplete) prophage regions. For the first time, the characteristics of the genome of this species were described and a pangenomic analysis was performed. The concept of the pangenome was used not only to establish the genetic repertoire of this species, but primarily to highlight the unique characteristics of L. hilgardii FLUB. The core of the genome of L. hilgardii is centered around genes related to the storage and processing of genetic information, as well as to carbohydrate and amino acid metabolism. Strains with such a genetic constitution can effectively adapt to environmental changes. L. hilgardii FLUB is distinguished by an extensive cluster of metabolic genes, arsenic detoxification genes, and unique surface layer proteins. Variants of MRS broth with ethanol (10–20%), glucose (2–25%), and fructose (2–24%) were prepared to test the strain’s growth preferences using Bioscreen C and the PYTHON script. L. hilgardii FLUB was found to be more resistant than a reference strain to high concentrations of alcohol (18%) and sugars (25%). It exhibited greater preference for fructose than glucose, which suggests it has a fructophilic nature. Comparative genomic analysis supported by experimental research imitating the conditions of alcoholic beverages confirmed the niche specialization of L. hilgardii FLUB to the mead environment.

This chapter presents the possibility of using cold plasma in sustainable food production. First, the concept of cold plasma was defined, the sources of plasma production and the mechanism of antimicrobial activity were characterized. Next, the use of cold plasma in the preservation of food products was described and explained its influence on the physicochemical properties of food products. The following section describes in detail the main areas in which cold plasma can be used for sustainable food production, including both the agricultural sector and the food industry. At the end of the chapter, attention was paid to the challenges for scientists to safely introduce cold plasma technology into commercial applications.

Atmospheric cold plasma (ACP) inactivation of Lentilactobacillus hilgardii was investigated. Bacteria were exposed to ACP dielectric barrier discharge with helium and oxygen as working gases for 5, 10, and 15 min. The innovative approach in our work for evaluation of bacterial survival was the use in addition to the classical plate culture method also flow cytometry which allowed the cells to be sorted and revealed different physiological states after the plasma treatment. Results showed total inhibition of bacterial growth after 10-min of ACP exposure. However, the analysis of flow cytometry demonstrated the presence of 14.4% of active cells 77.5% of cells in the mid-active state and 8.1% of dead cells after 10 min. In addition, some of the cells in the mid-active state showed the ability to grow again on culture medium, thus confirming the hypothesis of induction of VBNC state in L .hilgardii cells by cold plasma. In turn, atomic force microscopy (AFM) which was used to study morphological changes in L. hilgardii after plasma treatment at particular physiological states (active, mid-active, dead), showed that the surface roughness of the mid-active cell (2.70 ± 0.75 nm) was similar to that of the control sample (2.04 ± 0.55 nm). The lack of considerable changes on the cell surface additionally explains the effective cell resuscitation. To the best of our knowledge, AFM was used for the first time in this work to analyze cells which have been sorted into subpopulations after cold plasma treatment and this is the first work indicating the induction of VBNC state in L. hilgardii cells after exposure to cold plasma.

This paper demonstrates the role of curly kale fermentation by autochthonous lactic acid bacteria in the formation of bioactive derivatives of phenolic compounds which may have anticancer properties. LC-MS and HPLC analyses were performed and the content of polyphenols in curly kale before and during fermentation were determined. Bioactive properties of salicylic and gentisic acids were tested in experiments on tumor (HT29, SW620) and normal (CCD841CoTr) epithelial cell lines. In this study, gentisic and salicylic acids were thoroughly examined. Gentisic acid has not been detected in Brasssicaceae vegetables so far, and the presence of both these acids was noted in fermented curly kale for the first time. Our results indicated that both salicylic and gentisic acids decreased the number of HT29 cells simultaneously with reduction of the intercellular interactions between these cells. Moreover, immunomodulatory and anti-inflammatory activity of gentisic acid was observed.

In the present study, Lactobacillus plantarum strain was isolated and identified from spontaneous fermentation of Brassica oleracea var. capitata L. We used the Unweighted Pair Group Method with Arithmetic Mean Analysis (UPGMA) and Principal Component Analysis (PCA) to examine the variations in the functional properties of the isolates. Six functional traits were analyzed, i.e., viability at low pH, resistance to lysozyme and to SIF, auto- and coaggregation, and s-glucosidase activity. The present work is the first study in which the PCA and UPGMA statistical methods were used together to analyze data obtained from the same microbiological experiments. This provided information about the similarity of the examined isolates in terms of their functional traits. Additionally, the level of the analyzed functional traits within the particular groups of isolates was shown. The presented approach is the basis for choosing isolates that are most closely related to the reference strain isolated from pickled cabbage.

The present work is the first report on spontaneous fermentation of curly kale and characteristics of autochthonous lactic acid bacteria (LAB). Our results indicate that curly kale fermentation is the new possibility of the technological use of this vegetable. Bacteria representing ten different species were isolated from three phases of curly kale fermentation and identified by MALDI-TOF mass spectrometry and 16S rRNA gene sequencing. Among them, four species were identified as Lactobacillus spp. (Lb. plantarum 332, Lb. paraplantarum G2114, Lb. brevis R413, Lb. curvatus 154), two as Weissella spp. (W. hellenica 152, W. cibaria G44), two as Pediococcus spp. (P. pentosaceus 45AN, P. acidilactici 2211), one as Leuconostoc mesenteroides 153, and one as Lactococcus lactis 37BN. The functional properties of isolates, i.e. acid, NaCl and bile salt tolerance, enzyme activities, adhesion to hydrocarbons, and antibiotic resistance, were examined. Among the tested strains, Lb. plantarum 332, Lb. paraplantarum G2114, P. pentosaceus 2211, and Lb. brevis R413 exhibited the best hydrophobicity value and high tolerance to bile salts, NaCl, and low pH.

Microbiological contamination is a big challenge to the food industry, medicine, agriculture, and environmental protection. For this reason, scientists are constantly looking for alternative methods of decontamination, which ensure the effective elimination of unwanted biological agents. Cold plasma is a new technology, which due to its unique physical and chemical properties becomes a point of interest to a growing group of researchers. The previously conducted experiments confirm its effective action, e.g. in the disinfection of skin wounds, air, and sewage treatment, as well as in food preservation and decontamination. The reactive compounds present in the plasma: high-energy electrons, ionized atoms and molecules, and UV photons are the key factors that cause an effective reduction in the number of microorganisms. The mechanism and effectiveness of the cold plasma are complex and depend on the process parameters, environmental factors and the type and properties of the microorganisms that are to be killed. This review describes the current state of knowledge regarding the effectiveness of the cold plasma and characterizes its interaction with various groups of microorganisms based on the available literature data. Microbiological contamination is a big challenge to the food industry, medicine, agriculture, and environmental protection. For this reason, scientists are constantly looking for alternative methods of decontamination, which ensure the effective elimination of unwanted biological agents. Cold plasma is a new technology, which due to its unique physical and chemical properties becomes a point of interest to a growing group of researchers. The previously conducted experiments confirm its effective action, e.g. in the disinfection of skin wounds, air, and sewage treatment, as well as in food preservation and decontamination. The reactive compounds present in the plasma: high-energy electrons, ionized atoms and molecules, and UV photons are the key factors that cause an effective reduction in the number of microorganisms. The mechanism and effectiveness of the cold plasma are complex and depend on the process parameters, environmental factors and the type and properties of the microorganisms that are to be killed. This review describes the current state of knowledge regarding the effectiveness of the cold plasma and characterizes its interaction with various groups of microorganisms based on the available literature data.

Accumulation of toxic metal ions in food and water is nowadays a growing health-related problem. One detoxification method involves the use of microorganisms naturally inhabiting the gastrointestinal tract (GIT). The purpose of this study was to prove that lactic acid bacteria derived from the GIT are able to effectively remove Cd2+ from water solution. Seven strains of lactobacilli, out of 11 examined, showed tolerance to high concentrations of cadmium ions. The metal-removal efficiencies of these seven lactobacilli ranged from 6 to 138.4 μg/h mg. Among these bacteria, Lactobacillus gallinarum and Lactobacillus crispatus belonged to the highest (85%) Cd-removal efficiency class. An analysis of the zeta potential (ζ) indicated that the bacterial cell surface had a negative charge at the pH ranging from 3 to 10. The presence of carboxyl, amide, and phosphate groups was favorable for Cd2+ binding to the cell surface, which found confirmation in FTIR-ATR spectra. Elemental SEM/EDS analysis and TEM imaging not only confirmed the adsorption of Cd2+ on the cell envelope but also gave us a reason to suppose that Lb. crispatus accumulates metal ions inside the cell. Our findings open perspectives for further research on the new biological function of GIT lactobacilli as natural biosorbents.

This is the first report on the physicochemical properties of chitin obtained from larvae and imagoes of black soldier flies (Hermetia illucens). Scanning electron microscopy revealed differences in surface morphologies of the two types of chitin. The crystalline index values of chitins from adult flies and larvae were 24.9% and 35%, respectively. This is a trait that differentiates these biopolymers from chitins extracted from other sources described so far. X-ray diffraction patterns and IR spectroscopy revealed that both types of samples of chitin were in an α crystalline form. Also, the results of elemental analysis, thermal stabilities and FTIR spectroscopy of the chitins from larvae and adults of H. illucens were similar, which points to a general similarity in their physicochemical structure.

Multivariate statistical analysis was used to select potential starter cultures to produce fermented curly kale juice. The technological and functional properties of juices fermented using 10 autochthonous lactic acid bacteria isolated from spontaneously fermented curly kale were examined based on their capability of fast acidification while maintaining high numbers of viable cells, antioxidant activity, and antimicrobial effects against Gram positive and Gram negative bacteria. The outcome of the analysis of these parameters was used as input data for a principal component analysis (PCA) to select the most promising strains. The PCA showed that three strains Lactobacillus paraplantarum G2114 (GenBank ID: KY883562 ), Lb. plantarum 332 (GenBank ID: KY883559 ), and Pediococcus pentosaceus 2211 (GenBank ID: KY883563 ) showed a similarity to Lb. plantarum NRRL B-4496 used as a reference strain in terms of functional and technological properties and its ability to ferment vegetables. These strains were selected as starter culture for further fermentations. Samples of curly kale juice fermented with a multistrain formulation showed fast acidification with a high number of viable cells, high level of antioxidant activity and antimicrobial effects against some pathogenic bacteria. Furthermore, the multistrain formulation resulted in improved results in comparison with the control and autochthonous bacteria strains tested individually.

In the present study, a Lactobacillus plantarum FPL strain exhibiting fructophilic behavior has been isolated for the first time from honeydew. It is a probably syntrophic bacterium inhabiting the gastrointestinal tract of Coccus hesperidum L. and taking part in sugar metabolism. The promising growth characteristics and biochemical properties of Lb. plantarum FPL indicate that this may be a facultatively fructophilic species, whose properties are not associated with the loss of the alcohol/acetaldehyde dehydrogenase gene. The article attempts to classify the peculiar behavior of this strain by means of tests that are characteristic for FLAB as well as through a classic identification approach. In this study, we used a reference strain Lb. plantarum NRRL B-4496, which showed no fructophilic properties. With the FLAB group, the new strain shares the habit, such as a fructose-rich environment, the preference of this sugar for growth, and similar growth curves. However, it exceeds FLAB in terms of osmotolerance to high sugar content. The fructophilic Lb. plantarum FPL strain can proliferate and grow on a medium wherein the sugar concentration is 45 and 50% (w/v). Our findings indicate that honeydew can be a promising source of new fructophilic lactic acid bacteria. Electronic supplementary material The online version of this article (10.1007/s13213-018-1350-2) contains supplementary material, which is available to authorized users.

The aim of this study was to assess the removal of Cd(2+) and Al(3+) from aqueous solutions by an exopolysaccharide (EPS) from Lactobacillus rhamnosus E/N. The biosorption kinetics of EPS for Cd(2+) and Al(3+) were studied by equilibrium dialysis after incubation from 0 h to 58 h. The actual concentrations of Cd(2+) and Al(3+) in external solutions were measured by a fluorescence method after complexation with 8-hydroxyquinoline. Binding of metal ions by EPS was observed, with maximum adsorption after 0 to 24 and 48 to 58 h for Cd(2+) , and after 24 h for Al(3+) . The existing analytical method, involving the binding of metal ions was measured using a dialysis membrane was significantly improved in our study. This new method could be applied in future studies of this type. The results of this study demonstrate that EPS from Lb. rhamnosus E/N has a Cd(2+) and Al(3+) biosorption capacity. Surface adsorption of the metal ions at surface of EPS was confirmed through scanning electron microscopy. Fourier transform-infrared spectroscopy analysis suggests a role of the functional groups ─OH, C═O, and COO(-) from EPS in the binding of Cd(2+) and Al(3+) ions.

The present paper describes optimization of fermentation conditions in shaken flasks and scale-up of fermentor production up to 115 L. The response surface methodology (RSM) has been successfully applied in standardization of mutanase production by Trichoderma harzianum CCM F-340. The model was very well fitted to the experimental data and explained more than 96% of the whole variation of the response (adjusted R2 = 0.962). In order to confirm the adequacy of the regression model based on the experimental data, validation cultures were grown in conditions created through optimization. The highest enzyme activity (0.747 U/mL) was reached in shaken flask cultures on Mandels’ medium in a volume of 140 mL modified in terms of carbon (cell wall preparation from the polypore fungus Laetiporus sulphureus 8.08 g/L) and nitrogen (soybean peptone 1.38 g/L) sources, under culture conditions 30°C, pH 5.3, agitation 270 rpm. The scale-up of the culture in the bioreactors with a working volume of 5 and 115 L resulted in a slight decrease in the mutanase activity (0.734 and 0.682 U/mL, respectively). The validation experiment showed a 70.6% increase in the production of mutanase compared with the culture before optimization. The results proved that the cultures could be scaled-up successfully from shaken flasks to the bioreactor scale. Our results indicate that in optimal conditions, T. harzianum could be a highly effective extracellular mutanase source. This report is the first to deal with optimization of mutanase biosynthesis using a mathematical model and scale-up of enzyme production in controlled fermentors with a view to facilitate application thereof in industry.Keywords: Mutanase, Trichoderma harzianum, response surface methodology (RSM), bioreactors, submerged culture.African Journal of Biotechnology, Vol 13(25) 2538-2546

The presence of S-layer proteins in the cell envelope of Lactobacillus helveticus may be technologically important. S-layer proteins are the adhesion site for cell envelope proteinase, which forms the proteolytic pathway in bacteria. Eleven strains of L. helveticus were examined for the presence of S-layer proteins and slpH genes. S-layer proteins from six strains were identified and sequenced. Multiple alignments of the deduced amino acid sequences demonstrated a strong sequence conservation of all Slp studied. Transmission Electron Microscopy analysis of the cells revealed the typical cell wall architecture of the S-layer. This is the first report on characterisation of glycosylated S-layer proteins from different strains of L. helveticus. The amino acid composition, the secondary structure, and the physical properties of these proteins were found to be quite similar to those of S-layer proteins from other lactobacilli. However, PCR analysis revealed that five of the examined strains of L. helveticus did not have slpH genes. This finding suggests that S-layer protein genes cannot be considered as housekeeping genes and cannot be used as molecular markers for L. helveticus.

Genetic mechanisms of variation in erythromycin resistance in Lactobacillus rhamnosus strains

In this study sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) profiles were analysed and differences were confirmed by a unweighted pair group method with arithmetic average (UPGMA) analysis between bifidobacterial species, such as B. infanis ATCC1567, B. bifidum Bb-12, B. longum KN29, B. catenulatum KD14, and B. animalis BI30. Two dimensional electrophoresis separation profiles were compared, and the most characteristic spots were characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We propose proteins extracted from intact cells as an additional trait for bifidobacteria characterization, together with molecular techniques, which can be used to analyze bacterial protein polymorphism and to distinguish among species.

Response surface methodology (RSM) was employed to study the effects of various medium components on biomass production by Lactobacillus rhamnosus E/N. This strain is commonly used in the pharmaceutical and food industries due to its beneficial effect on the human gut and general health. The best medium composition derived from RSM regression was (in g/l) glucose 15.44, sodium pyruvate 3.92, meat extract 8.0, potassium phosphate 1.88, sodium acetate 4.7, and ammonium citrate 1.88. With this medium composition biomass production was 23 g/l of dry cell weight after 18 h of cultivation in bioreactor conditions, whereas on MRS the yield of biomass was 21 g/l of dry cell weight. The cost of 1 g of biomass obtained on MRS broth was calculated at the level of 0.44 € whereas on the new optimal medium it was 25% lower. It may be concluded then, that the new medium, being cheaper than the control MRS allows large scale commercial cultivation of the L. rhamnosus strain. This study is of relevance to food industry because the possibility to obtain high yield of bacterial biomass is necessary step in manufacturing of probiotic food.

This paper describes a simple and efficient method of isolation of a plullulanase type I from amylolytic lactic acid bacteria (ALAB). Extracellular pullulanase type I was purified from a cell-free culture supernatant of Lactococcus lactis IBB 500 by using ammonium sulfate fractionation and dialysis (instead of ultrafiltration), and ion-exchange chromatography with CM Sepharose FF followed by gel filtration chromatography with Sephadex G-150 as the final step. A final purification factor of 14.36 was achieved. The molecular mass of the enzyme was estimated as 73.9 kD. The optimum temperature for the enzyme activity was 45°C and the optimum pH was 4.5. Pullulanase activity was increased by addition Co(2+) and completely inhibited by Hg(2+). The enzyme activity was specifically directed toward α-1,6 glycosidic linkages of pullulan giving maltotriose units. Enzymatic hydrolysis of starch and amylose produced a mixture of maltose and maltotriose.

Aims: Response surface methodology (RSM) was used to optimize a protective medium for enhancing the viability of Lactobacillus rhamnosus E/N cells during lyophilization. Methods and Results: Spirulina, sucrose and lactulose were selected, on the basis of a Plackett-Burman factorial design, as important protectants having the following protective effects on cell viability: 102·025, 36·885 and −34·42, respectively. A full-factorial central composite design was applied to determine optimal levels of three used agents. Conclusion: The optimal protective medium composition was determined to be: Spirulina 1·304% (w/v), lactulose 5·48% (w/v), and sucrose 13·04% (w/v) (Polish Patent P-393189). The predictive value of cell viability in this medium was 89·619%, and experimental viability obtained during freeze-drying was 87·5%. Significance and Impact of the Study: In this study, Spirulina was used for the first time as the protective agent in freeze-drying medium, significantly increasing lactobacilli viability and giving synbiotic character of the final product.

The impact of five carbohydrate sources (glucose, maltose, galactose, sucrose, and lactose) on the chemical composition, structure, morphology, and physicochemical properties, as well as, viscosity of exopolysaccharides (EPSs) produced by Lactobacillus rhamnosus E/N was investigated. GLC-MS analysis and 2DNMR spectroscopy showed that the EPSs had the same primary structure independently of the carbon source used in the growth medium. The following EPS composition was elucidated: four rhamnose, two glucose, and one galactose residue with a pyruvate substituent. Molecular masses (M(w)) were determined by gel permeation chromatography, which revealed differences in M(w) distribution. EPS-Gal, EPS-Suc, and EPS-Lac showed heterogenic fractions of a high and low molecular weight, while EPS-Mal and EPS-Glc contained only a high-molecular-weight fraction. AFM microscopy revealed morphological differences in chain length, thickness, and branching. Differences in the Mw ratio and thickness of the polymer chain were correlated with high viscosity of EPS solutions. Our results indicate that a single bacterial strain, depending on the carbon source in the medium, can produce EPSs of different rheological properties.

The goal of this study was to identify moonlighting proteins in Lactobacillus helveticus that play an important role in adhesion and aggregation. The label-free method was used for identification and analysis of expression of cellular proteins. The analysis revealed the presence of eight moonlighting proteins in the cell envelope of Lb. helveticus. The tested strains mainly differed with respect to the presence of S-layer proteins and the level of expression of moonlighting proteins in Lb. helveticus strain T159. These surface proteins give the cell a hydrophobic character and play a role in specific interactions with intestinal epithelium cells and with other bacteria. In Lb. helveticus T159, the S-layer associated with moonlighting proteins could act as adherence factors, which was evidenced by the high capability of adhesion, auto- and coaggregation. The hydrophobicity, adhesion and aggregation abilities provide biological activities in food products and they are regarded as an important criterion for probiotic selection.

The aim of this study was to assess the removal of Cd(2+) and Al(3+) from aqueous solutions by an exopolysaccharide (EPS) from Lactobacillus rhamnosus E/N. The biosorption kinetics of EPS for Cd(2+) and Al(3+) were studied by equilibrium dialysis after incubation from 0 h to 58 h. The actual concentrations of Cd(2+) and Al(3+) in external solutions were measured by a fluorescence method after complexation with 8-hydroxyquinoline. Binding of metal ions by EPS was observed, with maximum adsorption after 0 to 24 and 48 to 58 h for Cd(2+) , and after 24 h for Al(3+) . The existing analytical method, involving the binding of metal ions was measured using a dialysis membrane was significantly improved in our study. This new method could be applied in future studies of this type. The results of this study demonstrate that EPS from Lb. rhamnosus E/N has a Cd(2+) and Al(3+) biosorption capacity. Surface adsorption of the metal ions at surface of EPS was confirmed through scanning electron microscopy. Fourier transform-infrared spectroscopy analysis suggests a role of the functional groups ─OH, C═O, and COO(-) from EPS in the binding of Cd(2+) and Al(3+) ions.

Lactobacillus rhamnosus E/N is a probiotic bacterium, which synthesizes exopolysaccharides (EPS) with significant bifidogenic and antioxidant activities. The sugar composition of the EPSs produced depended on carbohydrates used as a carbon source in the growth media. Five Bifidobacterium strains were tested in vitro for their ability to utilize all the EPSs studied. The highest bifidogenic activity was revealed by EPSs obtained from Lactobacillus cultures supplemented with Gal, Lac, and Mal as the only carbon source, while significant antioxidant effects were observed in EPSs isolated from growth media enriched with galactose, lactose, and sucrose.

An extracellular alpha-amylase from Lactococcus lactis IBB500 was purified and characterized. The optimum conditions for the enzyme activity were pH 4.5, temperature of 35 degrees C, enzyme molecular mass of 121 kDa. The genome analysis and a plasmid curing experiment indicated that amy+ genes were located in a plasmid of 30 kb. An analysis of phylogenetic relationships strongly supported a hypothesis of horizontal gene transfer. A strong homology was found for the peptides with the sequence of alpha-amylases from Ralstonia pikettii and Ralstonia solanacearum. The protein of alpha-amylase activity purified in this study is the first one described for the Lactococcus lactis species, and this paper is the first report on Lactococcus lactis strain as a microorganism belonging to amylolytic lactic acid bacteria (ALAB).

The central composite design was developed to search for an optimal medium for the growth of Lactobacillus rhamnosus OXY. The effect of various media components, such as carbon sources, simple and complex nitrogen sources, mineral agents, and growth factors (vitamins B, amino acids) was examined. The first-order model based on Plackett-Burman design showed that glucose, sodium pyruvate, meat extract and mineral salts significantly influenced the growth of the examined bacteria. The second-order polynomial regression confirmed that maximum biomass production could be achieved by the combination of glucose (12.38 g/l), sodium pyruvate (3.15 g/l), meat extract (4.08 g/l), potassium phosphate (1.46 g/l), sodium acetate (3.65 g/l) and ammonium citrate (1.46 g/l). The validation of the predicted model carried out in bioreactor conditions confirmed the usefulness of the new medium for the culture of L. rhamnosus OXY in large scale. The optimal medium makes the culture of the probiotic bacterium L. rhamnosus OXY more cost effective.

Different methods were used to detect antibacterial activity of Lactobacillus rhamnosus against some food spoilage microorganisms. This study demonstrates that in comparison with culture supernatant and sonificated cells of L. rhamnosus the strongest antimicrobial activity was observed when live test culture was used. It can be suggested that the best results were seen with two methods: the dual culture overlay assay and the agar slab method. From comparison between methodologies it can be concluded that the agar slab technique gives the most reliable and reproducible results.

The aim of this study was to analyze the cell envelope components and surface properties of two phenotypes of Lactobacillus rhamnosus isolated from the human gastrointestinal tract. The ability of the bacteria to adhere to human intestinal cells and to aggregate with other bacteria was determined. L. rhamnosus strains E/N and PEN differed with regard to the presence of exopolysaccharides (EPS) and specific surface proteins. Transmission electron microscopy showed differences in the structure of the outer cell surface of the strains tested. Bacterial surface properties were analyzed by Fourier transform infrared spectroscopy, fatty acid methyl esters and hydrophobicity assays. Aggregation capacity and adhesion of the tested strains to the human colon adenocarcinoma cell line HT29 was determined. The results indicated a high adhesion and aggregation ability of L. rhamnosus PEN, which possessed specific surface proteins, had a unique fatty acid content, and did not synthesize EPS. Adherence of L. rhamnosus was dependent on specific interactions and was promoted by surface proteins (42–114 kDa) and specific fatty acids. Polysaccharides likely hindered bacterial adhesion and aggregation by masking protein receptors. This study provides information on the cell envelope constituents of lactobacilli that influence bacterial aggregation and adhesion to intestinal cells. This knowledge will help to understand better their specific contribution in commensal–host interactions and adaptation to this ecological niche.