Magdalena Polak-Berecka

Physicochemical characterization of exopolysaccharides produced by Lactobacillus rhamnosus on various carbon sources

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 effect of moonlighting proteins on the adhesion and aggregation ability of Lactobacillus helveticus.

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.

Biosorption of Al(+3) and Cd(+2) by an exopolysaccharide from Lactobacillus rhamnosus.

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.

PURIFICATION AND CHARACTERIZATION OF PULLULANASE FROM Lactococcus lactis

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 Co2+ and completely inhibited by Hg2+. 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.

The first report of the physicochemical structure of chitin isolated from Hermetia illucens.

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.

Spirulina enhances the viability of Lactobacillus rhamnosus E/N after freeze‐drying in a protective medium of sucrose and lactulose

Aims:  Response surface methodology (RSM) was used to optimize a protective medium for enhancing the viability of Lactobacillus rhamnosus E/N cells during lyophilization.

The plackett-burman design in optimization of media components for biomass production of Lactobacillus rhamnosus OXY.

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.

Genetic mechanisms of variation in erythromycin resistance in Lactobacillus rhamnosus strains.

Genetic mechanisms of variation in erythromycin resistance in Lactobacillus rhamnosus strains

Composition of lactic acid bacteria during spontaneous curly kale (Brassica oleracea var. sabellica) fermentation

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.

Functional traits of Lactobacillus plantarum from fermented Brassica oleracea var. capitata L. in view of multivariate statistical analysis

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 ß-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.