Bacterial Exopolysaccharides Research Articles

OBTAINING OF BACTERIAL POLYSACCHARIDES

The aim of this work was to develop a technology for obtaining bacterial exopolysaccharides. The problem was solved by using the bacterial strain Halovibrio variabilis UzAS3 as a polysaccharide producer. The optimal parameters of bacterial cultivation have been determined. A technological scheme for obtaining bacterial polysaccharides has been developed, which includes such stages as the cultivation of bacteria, the separation of bacterial biomass from the culture liquid, desalting, precipitation, washing and drying. The obtained results show that bacteria have a high potential for the yield of bacterial exopolysaccharides (7.1-9.5 mg/ml).

Formation and ripening of alginate-like exopolymer gel layers during and after membrane filtration

The properties of biofilm EPS are determined by the multiple interactions between its constituents and the surrounding environment. Because of the high complexity of biofilm EPS, its constituents' characterisation is still far from thorough, and identification of these interactions cannot be done yet. Therefore, we use gels of bacterial alginate-like exopolysaccharides (ALEs) as a model component for biofilm EPS in this work. These gels have been examined for their cohesive properties as a function of CaCl2 and KCl concentration. Hereto, ALE gel layers were formed on membranes by dead-end filtration of ALE solutions. Accumulation of the cations Ca2+ and K+ in the gels could be well predicted from a Donnan equilibrium model based on the fixed negative charges in the ALE. This suggests that there is no specific binding of Ca2+ to the ALE and that on the time scale of the experiments, the Ca2+ ions can distribute freely over the gel and the surrounding solution. The concentration of fixed negative charges in the ALE was estimated around 1 mmol/g VSS (volatile suspended solids, organic mass) from the Donnan equilibrium. Moreover, an accumulation of H+ was predicted. Gels with more CaCl2 in the supernatant were more compact and bore a higher osmotic pressure than those with less CaCl2, revealing the role of Ca2+ ions in the network crosslinking. It is hypothesised that this mechanism later transitions into a rearrangement of the ALE molecules, which eventually leads to a fibrous network structure with large voids.

Yeast exopolysaccharides and their physiological functions.

Mounting evidence indicated the capability of various microorganisms in biosynthesis of exopolysaccharides (EPSs). A wide range of evidence extensively investigated the ability of bacterial species for EPS synthesis and their favorable effects, so little is known regarding yeast species. Many factors like composition of growth media and fermentation conditions are related to the structural and physical properties of EPSs. The EPS protects the producer yeast strain against extreme environment. Researchers proposed that yeast EPSs have priority over bacterial EPSs because of high yields of EPS biosynthesis and easy separation methods from growth media. Besides, they have drawn increasing attention due to their interesting biological activities, food, pharmaceutical, and cosmetics applications. Although a limited number of studies exist, this review aims to highlight the EPS structure and various applications of known yeast species in detail.

Effect of electrostatic interactions and complexes formation between nisin and bacterial exopolysaccharides on nisin antimicrobial efficacy

This study aims to describe interactions that may occur between nisin and bacterial exopolysaccharides (EPS) when they are present together in a model solution. For this purpose, two EPS, namely EPS-GM and EPS-BMS, produced by potential probiotic Bacillus tequilensis-GM and Leuconostoc citreum-BMS, respectively, were used. Results showed that maximal complexation occurred at pH 5.0 between anionic EPS (1.5 g/L) and cationic nisin (1 g/L) mainly through electrostatic interactions. Results provided by scanning electron microscopy, epifluorescence microscopy and turbidity measurements showed that EPS-GM was able to form bigger and higher amounts of complexes with nisin. The nisin antimicrobial activity was interestingly not affected and even improved after its complexation with EPS suggesting that these latter may act as protective agents of the nisin antimicrobial activity when applied in foods. The results of this study should help food technologists to optimize the physicochemical conditions for optimal formulation of food products containing EPS.

Biomedical Applications of Bacterial Exopolysaccharides: A Review.

Bacterial exopolysaccharides (EPSs) are an essential group of compounds secreted by bacteria. These versatile EPSs are utilized individually or in combination with different materials for a broad range of biomedical field functions. The various applications can be explained by the vast number of derivatives with useful properties that can be controlled. This review offers insight on the current research trend of nine commonly used EPSs, their biosynthesis pathways, their characteristics, and the biomedical applications of these relevant bioproducts.

Biosynthesis of exopolysaccharides by soil bacteria <i>Paenibacillus mucilaginosus</i> on a nutrient molasses medium

Bacterial exopolysaccharides exhibiting water-retaining and cementing properties play an essential role in such processes as the formation and stabilization of soil aggregates, regulation of nutrients and water flow through plant roots, as well as reduction of salt stress in plants. When bacteria producing exopolysaccharides are introduced into a soil, plants become more resistant to water stress due to improved structure and significant accumulation of proline, sugars and free amino acids under the conditions of water deficiency. In this work, we set out to determine the efficiency of exopolysaccharide biosynthesis by the soil bacteria Paenibacillus mucilaginosus when cultivated on a nutrient molasses medium. The bacterial strains P. mucilaginosus 560 and 574 provided by the Departmental collection of non-pathogenic microorganisms for agricultural purposes (All-Russia Research Institute for Agricultural Microbiology, St. Petersburg) were used as exopolysaccharide producers. In order to determine the influence of cultivation conditions on the synthesis of exopolysaccharides by P. mucilaginosus, the one-factor-at-a-time (OFAT) method was used. On the basis of the obtained results, we determined the most effective producer of exopolysaccharides, the age and dose of the inoculum, the optimal values of the cultivation temperature and medium pH, the source of nitrogen and its concentration, as well as the aeration conditions affecting exopolysaccharide biosynthesis and the growth of the selected bacterial strain P. mucilaginosus. It was established that the P. mucilaginosus 574 strain is the most effective producer of exopolysaccharides. It was shown that the biosynthesis of exopolysaccharides is most effective and economical upon the cultivation of P. mucilaginosus 574 on a nutrient medium containing 2% molasses without addition of minerals and nitrogen. The maximum amount of exopolysaccharides can reach 9.55 g/L on a nutrient medium containing 2% molasses at a volume of 50 ml with the addition of 0.1% corn extract as an inducer of exopolysaccharide synthesis at a cultivation temperature of 30±1 °C, a medium pH of 6.0±0.2 with the introduction of 5% inoculum after 24 hours of inoculation. The obtained research results are recommended for use when creating new technologies for the production of microbiological fertilizers.

Bacterial Exopolysaccharides and Their Applications: A Short Review

Extracellular polymer substances (EPS) are metabolic products of bacteria which accumulate on the cell surface and serve as a protective layer against harsh environmental condition. EPS also serve as a source of energy during the starvation of bacteria. The major structural component of EPS is polysaccharide, commonly known as exopolysaccharide, is either neutral or polyanionic in nature. Polysaccharides in EPS found different in sugar compositions and moieties, chain length, repeating units and substitutions in the side chains. Recently, the usefulness of bacterial exopolysaccharides, has led to discovery by advance technology are found to numerous industrial and medical applications. The properties of exopolysaccharides have changed the industrial and medical sectors due to their various applications and prospects. These applications have been comprehensive in areas such as nutraceutical functional food, cosmetics, pharmacological, herbicides and insecticides, while prospects include uses as antithrombotic, immunomodulator, anticancer and as bio flocculants. Go through this article to know different types of bacterial exopolysaccharides, and their different functions in human beings.

Biological and microbiological activities of isolated Enterobacter sp. ACD2 exopolysaccharides from Tabuk region of Saudi Arabia

Several forms of bacterial exopolysaccharides (EPSs) have been reported as having industrial and medical benefits. Previously, our group had achieved a high yield of EPS production (8.6 gm/L) from Enterobacter sp. isolated from the marine environment of Haqel Beach, in the Tabuk region of Saudi Arabia, by optimizing its culture conditions. Then, the strain selected was identified using conventional methods and confirmed by molecular characterization that used 16S Ribosomal RNA (16S rRNA) gene sequencing. In the present study, we determined the composition of this EPS and evaluated its anticoagulation, fibrinolytic, antimicrobial, and prebiotic activities. We determined the monosaccharide moieties of this EPS using acid hydrolysis by qualitative and quantitative paper chromatography (PC). The monosaccharide composition of this EPS was almost 25% glucose, 25% galactose, 40% fucose, in addition to 10% uronic acid, with traces of fructose. Sulfation modification was applied to the EPS produced. The biological and microbiological (anticoagulation, fibrinolytic, antimicrobial, and prebiotic) activities of both the native and sulfated EPS were investigated and both exhibited higher antibacterial activity against Staphyllococcus aureus and Escherichia coli, although neither form displayed antifungal activity against Candida albicans. The minimum inhibitory concentration (MIC) of the native and sulfated EPS that sustained the highest bacterial inhibition zone was 15 mg/dl. No prebiotic activities were recorded for either EPS. In conclusion, the biopolymer produced has significant biological activities that require further adaptation for practical and industrial applications.

Benefits of bacteria-derived exopolysaccharides on gastrointestinal microbiota, immunity and health

The gastrointestinal tract (GIT) is colonized by a broad spectrum of microorganism, of which bacteria are especially recognized for the health effects they exert. However insight in the effector molecules on the bacteria responsible are still largely lacking. Recently, bacterial exopolysaccharides (EPS), which are glycan structures on the bacterial cell wall, have gained considerable attention for their effects on human health. EPS are found to act as polymers that directly influence bacterial interactions with their host, such as bacterial adhesion to the GIT epithelium, their influence on immune responses, and the intestinal microbiota. These effects are highly dependent on the type of EPS structures and might be related to specific monosaccharides or linkages in EPS. Here, current knowledge on structure-specific EPS functions is reviewed in view of possible use of EPS to stimulate intestinal health and the formulation of dietary EPS mixtures or the possible creation of new EPS structures.

Lactic acid bacterial exopolysaccharides strongly bind histamine and can potentially be used to remove histamine contamination in food.

The symptoms of foodborne histamine poisoning are similar to those of IgE-mediated food allergies. In this study, we investigated the histamine-binding capacity of lactic acid bacteria (LAB) strains as potential preventive agents against histamine poisoning. Histamine biosorption capacity was determined for 16 LAB strains. Leuconostoc mesenteroides TOKAI 51 m, Lactobacillus paracasei TOKAI 65 m, Lactobacillus plantarum TOKAI 111 m and Pediococcus pentosaceus TOKAI 759 m showed especially high biosorption rates and reached saturation within 30 min. Adsorption isotherms showed better conformance to the Freundlich model than to the Langmuir model. Analyses after heat, periodic acid and guanidine hydrochloride treatments suggested that histamine was bound to the bacterial cell surface. HPLC analysis revealed that exopolysaccharides produced by Lact. paracasei TOKAI 65 m strongly bound to histamine. In the detachment test with 1 mol l-1 HCl solution, the dissociation rate of histamine for Lact. paracasei TOKAI 65 m was <10 %. This strain is presumably a suitable candidate for use against histamine poisoning.

The second messenger c-di-AMP mediates bacterial exopolysaccharide biosynthesis: a review.

Cyclic dimeric adenosine 3'-5'-monophosphate (c-di-AMP) is a recently discovered nucleotide messenger in bacteria. It plays an important role in signaling, transcription, and cell physiology, such as in bacterial growth, potassium transport, fatty acid synthesis, the metabolic balance of cell wall components, and biofilm formation. Exopolysaccharides (EPSs) have distinct physico-chemical properties and diverse bioactivities including antibacterial, hypolipidemic, and antioxidative activities, and they are widely used in the food, pharmaceutical, and cosmetic industries. Although c-di-AMP has been demonstrated to regulate the biosynthesis of bacterial EPSs, only a single c-di-AMP receptor, CabpA, has been identified in EPS synthesis. With the aim of describing current understanding of the regulation of microbial EPSs, this review summarizes c-di-AMP biosynthesis and degradation as well as the mechanism through which c-di-AMP regulates bacterial EPSs.

Water Conservation and Plant Survival Strategies of Rhizobacteria under Drought Stress

Drylands are stressful environment for plants growth and production. Plant growth-promoting rhizobacteria (PGPR) acts as a rampart against the adverse impacts of drought stress in drylands and enhances plant growth and is helpful in agricultural sustainability. PGPR improves drought tolerance by implicating physio-chemical modifications called rhizobacterial-induced drought endurance and resilience (RIDER). The RIDER response includes; alterations of phytohormonal levels, metabolic adjustments, production of bacterial exopolysaccharides (EPS), biofilm formation, and antioxidant resistance, including the accumulation of many suitable organic solutes such as carbohydrates, amino acids, and polyamines. Modulation of moisture status by these PGPRs is one of the primary mechanisms regulating plant growth, but studies on their effect on plant survival are scarce in sandy/desert soil. It was found that inoculated plants showed high tolerance to water-deficient conditions by delaying dehydration and maintaining the plant’s water status at an optimal level. PGPR inoculated plants had a high recovery rate after rewatering interms of similar biomass at flowering compared to non-stressed plants. These rhizobacteria enhance plant tolerance and also elicit induced systemic resistance of plants to water scarcity. PGPR also improves the root growth and root architecture, thereby improving nutrient and water uptake. PGPR promoted accumulation of stress-responsive plant metabolites such as amino acids, sugars, and sugar alcohols. These metabolites play a substantial role in regulating plant growth and development and strengthen the plant’s defensive system against various biotic and abiotic stresses, in particular drought stress.

Isolation and characterization of indigenous Weissella confusa for in situ bacterial exopolysaccharides (EPS) production in chickpea sourdough

Legume-based sourdough represents a potential ingredient for the manufacture of novel baked products. However, the lack of gluten of legume flours can restrict their use due to their poor technological properties. To overcome such issue, the in situ production of bacterial exopolysaccharides (EPS) during fermentation has been proposed. In this study, an EPS-producing lactic acid bacteria for in situ production in chickpea sourdough was isolated. After several backsloppings of the spontaneously fermented chickpea flour dough, a dominant strain of Weissella confusa was isolated and identified. W. confusa Ck15 was able to produce linear dextran with 2.6% α-(1 → 3) linked branches, from sucrose. Temperature of 30 °C, dough yield of 333, and 2% of sucrose addition were used to produce fermented chickpea sourdoughs. The acidification and rheology of the sourdoughs inoculated with W. confusa Ck15, Leuconostoc pseudomesenteroides DSM 20193, as positive control, and Lactobacillus plantarum F8, as negative control, were compared. The in situ dextran production by W. confusa Ck15 fermentation led to the highest viscosity increase (5.90 Pa·s) and the highest EPS percentage in the doughs (1.49%), compared to the other doughs. The in situ dextran production represents a potential approach for improving the use of legume flour in bakery products; overall, this experiment represents a first step for the exploitation of microbial EPS for setting up a baking process for chickpea based product.

Biosorption of Heavy Metals by the Bacterial Exopolysaccharide FucoPol

Despite the efforts for minimizing the usage of heavy metals, anthropogenic activities still generate high amounts of wastewater containing these contaminants that cause significant health and environmental problems. Given the drawbacks of the conventional physical and chemical methods currently used, natural biosorbents (microbial cells or their products) arise as promising environmentally friendly alternatives. In this study, the binding efficiency of the polysaccharide secreted by Enterobacter A47, FucoPol, towards lead (Pb2+), cobalt (Co2+), copper (Cu2+) and zinc (Zn2+) cations was demonstrated. FucoPol revealed a higher performance for the biosorption of Pb2+, with a maximum overall metal removal of 93.9 ± 5.3% and a specific metal uptake of 41.1 ± 2.3 mg/gEPS, from a Pb2+ solution with an initial concentration of 10 mg/L, by a 5 g/L FucoPol solution. The overall metal removal decreased considerably (≤31.3 ± 1.6%) for higher Pb2+ concentrations (48 and 100 mg/L) probably due to the saturation of FucoPol’s binding sites. Pb2+ removal was also less efficient (66.0 ± 8.2%) when a higher FucoPol concentration (10 g/L) was tested. Pb2+ removal efficiency of FucoPol was maximized at pH 4.3, however, it was affected by lower pH values (2.5–3.3). Moreover, the FucoPol’s sorption performance was unaffected (overall metal removal: 91.6–93.9%) in the temperature range of 5–40 °C. These findings demonstrate FucoPol’s great potential for utilization as a biodegradable and safe biosorbent for treating waters and wastewaters contaminated with Pb2+.

Overproduction of Exopolysaccharide Colanic Acid by Escherichia coli by Strain Engineering and Media Optimization.

Colanic acid (CA) is one of the major bacterial exopolysaccharides. Due to its biological activities, CA has a significant commercial value. However, the cultivation conditions have not been optimized for the large-scale production of CA. Here, we constructed a CA-overproducing Escherichia coli strain (ΔwaaF) and statistically optimized its culture media for maximum CA production. Glucose and tryptone were found the optimal carbon and nitrogen sources, respectively. Fractional factorial design indicated tryptone and Na2HPO4 as the critical nutrients for CA production. Through further optimization, we achieved a maximum CA production of 1910.0 mg/L, which is approximately 12-fold higher than the amount obtained using the non-optimized medium initially used. The predicted value of CA production was comparable with experimental value (2052.8 mg/L) under the optimized conditions. This study constitutes a successful demonstration of media optimization for increased CA production, and paves the way for future research for achieving large-scale CA production.

Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-Resistant Acinetobacter baumannii.

Acinetobacter baumannii is a leading cause of healthcare-associated infections worldwide. Its various intrinsic and acquired mechanisms of antibiotic resistance make the therapeutic challenge even more serious. One of the promising alternative treatments that is increasingly highlighted is phage therapy, the therapeutic use of bacteriophages to treat bacterial infections. Two phages active against nosocomial carbapenem-resistant A. baumannii strain 6077/12, vB_AbaM_ISTD, and vB_AbaM_NOVI, were isolated from Belgrade wastewaters, purified, and concentrated using CsCl gradient ultracentrifugation. The phages were screened against 103 clinical isolates of A. baumannii from a laboratory collection and characterized based on plaque and virion morphology, host range, adsorption rate, and one-step growth curve. Given that phage ISTD showed a broader host range, better adsorption rate, shorter latent period, and larger burst size, its ability to lyse planktonic and biofilm-embedded cells was tested in detail. Phage ISTD yielded a 3.5- and 2-log reduction in planktonic and biofilm-associated viable bacterial cell count, respectively, but the effect was time-dependent. Both phages produced growing turbid halos around plaques indicating the synthesis of depolymerases, enzymes capable of degrading bacterial exopolysaccharides. Halos tested positive for presence of phages in the proximity of the plaque, but not further from the plaque, which indicates that the observed halo enlargement is a consequence of enzyme diffusion through the agar, independently of the phages. This notion was also supported by the growing halos induced by phage preparations applied on pregrown bacterial lawns, indicating that depolymerizing effect was achieved also on non-dividing sensitive cells. Overall, good rates of growth, fast adsorption rate, broad host range, and high depolymerizing activity, as well as antibacterial effectiveness against planktonic and biofilm-associated bacteria, make these phages good candidates for potential application in combating A. baumannii infections.

Human apoptosis antibody array-membranes studying the apoptotic effect of marine bacterial exopolysaccharides in HepG2 cells.

Hepatocellular carcinoma (HCC) is considered as the third leading cause of cancer-related deaths, in spite of great advances in its treatment. The carbohydrate polymers, exopolysaccharides (EPSs), showed anticancer activity in diverse cancers. The purpose of this study is to investigate a panel of 43 apoptotic proteins to assess the possible apoptotic induction effect of bacterial EPSs showing promising cytotoxic effects in HepG2 cells in our previous study, in an attempt to introduce exopolysaccharides as new source for cancer treatment. Apoptosis-related proteins panel were examined through the analysis of Human Apoptosis Antibody Array-Membrane (43 targets). EPS-6 induces apoptosis through upregulation of different pro-apoptotic proteins as cytochrome C (9.52 fold) and tumor necrosis factor-related apoptosis-inducing ligand receptor (TRAIL-R1) (153.49 fold). EPS-RS induces apoptosis through up regulation of second mitochondria-derived activator of caspases (SMAC) (15.75 fold) and the six insulin-like growth factors binding proteins (IGFBP-1 through - 6) (76.81 fold, 7.68 fold, 55.15 fold, 4.9 × 107 fold, 29.69 fold, and 28.92 fold), respectively. Our results suggested that EPS-6 and EPS-RS could be considered as promising agents in hepatocellular carcinoma treatment.

Assembly of Bacterial Capsular Polysaccharides and Exopolysaccharides.

Polysaccharides are dominant features of most bacterial surfaces and are displayed in different formats. Many bacteria produce abundant long-chain capsular polysaccharides, which can maintain a strong association and form a capsule structure enveloping the cell and/or take the form of exopolysaccharides that are mostly secreted into the immediate environment. These polymers afford the producing bacteria protection from a wide range of physical, chemical, and biological stresses, support biofilms, and play critical roles in interactions between bacteria and their immediate environments. Their biological and physical properties also drive a variety of industrial and biomedical applications. Despite the immense variation in capsular polysaccharide and exopolysaccharide structures, patterns are evident in strategies used for their assembly and export. This review describes recent advances in understanding those strategies, based on a wealth of biochemical investigations of select prototypes, supported by complementary insight from expanding structural biology initiatives. This provides a framework to identify and distinguish new systems emanating from genomic studies.

Carvacrol oil inhibits biofilm formation and exopolysaccharide production of Enterobacter cloacae

This study aimed to analyze the effect of carvacrol oil on biofilm formation of Enterobacter cloacae. The minimum inhibitory concentration of carvacrol oil against E. cloacae was 256 μg/mL. The biofilm formation was inhibited by carvacrol oil at sub-inhibitory concentrations of 64 and 128 μg/mL. The bacterial cell motilities and biofilm exopolysaccharide (EPS) production were both inhibited by carvacrol oil. Correlation analysis indicated that the inhibition of biofilms in mature stages was related to the decrease of EPS production. The biofilm thickness and excretion of extrapolymeric matrix were reduced by carvacrol oil, as shown by scanning electron microscopy and confocal laser scanning microscopy analyses. Transcriptional analysis showed that carvacrol oil down-regulated 24 of 29 selected genes, including curli fimbriae genes (csgABCEFG) and colonic acid polysaccharide genes (wcaABCDEFKLM, wza, wzb, wzc, gmd, manB, manC) which are required for biofilm formation of E. cloacae. In addition, different antimicrobial agents, including 1% lactic acid, 1% phenyllactic acid, and 1024 μg/mL of carvacrol oil considerably inactivated the bacterial cells in carvacrol-treated biofilms. Hence, carvacrol oil effectively inhibited biofilm formation and EPS production of E. cloacae, which is useful for further inactivating E. cloacae biofilms by other antimicrobial agents.

Demonstration of the cryoprotective properties of the fucose-containing polysaccharide FucoPol

We report the cryoprotective potential of FucoPol, a fucose-containing bacterial exopolysaccharide produced by Enterobacter A47. In vitro cryopreservation assays of Vero, Saos-2, HFFF2 and C2C12 cell lines exposed to a validated non-cytotoxic 2.5 mg/mL FucoPol concentration demonstrated a consistent post-thaw metabolic viability increase. Calorimetric analysis showed a non-colligative, FucoPol concentration-dependent increase of the freezing point (Tf), with minimal change in melting point (Tm). Freezing point variation was corroborated by Polarized Optical Microscopy studies, also showing a reduction of ice crystal dimensions. Its proven shear-thinning behaviour and polyanionicity favour interactivity between the polysaccharide and the water-ice interface, resulting in ice growth inhibition. These findings demonstrate FucoPol’s high promise as a bio-based, biodegradable approach to be implemented into cryopreservation formulations.