Exopolysaccharides from Marine Bacteria

Microbial polysaccharides represent a class of important products of growing interest for many sectors of indus- try. In recent years, there has been a growing interest in isolating new exopolysaccharides (EPSs)-producing bacteria from marine environments, particularly from various extreme marine environments. Many new marine microbial EPSs with novel chemical compositions, properties and structures have been found to have potential applications in fields such as adhesives, textiles, Pharmaceuticals and medicine for anti-cancer, food additives, oil recovery and metal removal in mining and indus- trial waste treatments, etc This paper gives a brief summary of the information about the EPSs produced by marine bacteria, including their chemical compositions, properties and structures, together with their potential applications in industry.

A New Pullulan-Producing Yeast and Medium Optimization for Its Exopolysaccharide Production

Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pullulan was produced by Rhodotorula bacarum. The optimal medium (g L -1) for pullulan production by this strain was 80 glucose, 20 soybean cake hydrolysate, 5 K 2HPO 4, 1 NaCl, 0.2 MgSO 4·7H 2O, 0.6 (NH 4) 2SO 4, pH 7.0. Under this condition, 54 g L -1 pullulan was produced within 60 h at 30 ℃. Pullulan is a better starting material for producing marine prodrugs.

Production,characterization and biological activities of acidic exopolysaccharide from marine Bacillus amyloliquefaciens 3MS 2017

Objective: To evaluate in-vitro antioxidant, anti-inflammatory and antitumor abilities against human breast adenocarcinoma(MCF7) and human prostate cancer(PC3) as well as the suppressor effect of bacterial exopolysaccharide(BAEPS) on Ehrlich ascites carcinoma(EAC). Methods: In-vitro antioxidants characters of BAEPS were determined using various methods, while anti-inflammatory activity was estimated against cyclooxygenase(COX-1 and COX-2). In-vitro study, anticancer against MCF7 and PC3 were assessed by the mitochondrial dependent reduction of yellow MTT. In in-vivo study against EAC progression, mice were inoculated with EAC cells and then were orally administered BAEPS at 200 mg/kg after 24 h(equals to 0.10 of determined LD50)/10 d. Results: BAEPS was acidic exopolysaccharide contained uronic acid(12.3%) and sulfate(22.8%) with constitution of glucose, galactose and glucuronic acid in a molar ratio1.6: 1.0: 0.9, respectively, with a molecular mass of 3.76伊104 g/mol. BAEPS appeared potent antioxidant characters as free radical scavenging, oxygen reactive species scavenging and metal chelation, while its reducing power was low. BAEPS showed selective anti-inflammatory activity against COX-2 than COX-1, COX-2 selective. BAEPS exhibited potent and selective effect to breast cell cancer MCF7, the death percentage was 65.20% with IC_(50)=70 μg/m L and IC_(90)=127.40 μg/m L. BAEPS decreased counted viable EAC cells and induced non-viable cells. BAEPS improved all assessed hematological parameters. These improvements were reflected in the increasing median survival time and significant increment(P<0.05) in life span. Conclusions: BAEPS has anti-tumor activity with a good margin of safety. The anti-tumor activity of BAEPS may be due to its content from sulfated groups and uronic acids and they have antioxidant and anti-inflammatory properties.

Isolation, Purification, and Characterization of Exopolysaccharide Produced by Leuconostoc Citreum N21 from Dried Milk Cake

A strain with high production of exopolysaccharide(EPS) was isolated from dried milk cake(a traditional fermented food from Inner Mongolia). The strain was called N21 and later identified as Leuconostoc citreum( Leu. citreum). The strain was cultured in Man-Rogosa-Sharpe medium containing 50 g/L of sucrose for 48 h at 30 °C and the EPS purified, with a yield of 24.5 g/L. An average molecular weight of 6.07 × 10~6 g/mol was determined by high-performance size-exclusion chromatography. The structure of the purified EPS was investigated through gas chromatography, ~1H and ^(13)C nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. The results demonstrated a polysaccharide composed of D-glucopyranose units in a linear chain with consecutive α(1 → 6) linkages. No branching was found in the structure of the exopolysaccharide. The purified EPS showed high water solubility and emulsibility. Based on the thermogravimetric curve, the degradation temperature of the EPS was 308.47 °C, which suggested that the dextran in the study exhibited high thermal stability. The results indicated that Leu. citreum N21 could be widely used to produce linear EPS and that the EPS has potential applications in food science and processing as a food additive.

Optimization of medium components using response surface methodology (RSM) for mycelium biomass and exopolysaccharide production by <i>Lentinus squarrosulus</i>

The interaction between sucrose, yeast extract and initial pH was investigated to optimize critical medium components for mycelium biomass and production of exopolysaccharide (EPS) of Lentinus squarrosulus using Response Surface Methodology (RSM). A central composite design (CCD) was applied and a polynomial regression model with quadratic term was used to analyse the experimental data using analysis of variance (ANOVA). ANOVA analysis showed that the model was very significant (p Lentinus squarrosulus are as follows: sucrose concentration 114.61 g/L, yeast extract 1.62 g/L and initial pH of 5.81;sucrose concentration 115.8 g/L, yeast extract of 3.39 g/L and initial pH of 6.44 respectively.

Biosorption of Metal Ions by Exopolysaccharide Produced by <i>Azotobacter chroococcum</i>XU1

This paper deals with adsorption of Pb and Hg on the exopolysaccharide produced by Azotobacter chroococcum XU1. The adsorption of metal ions was significantly affected by the pH of metal solutions (Pb(NO3)2 and Hg(NO3)2), metal ions concentration and exopolysaccharide concentrations. At optimum pH the uptakes of the metals were 40.48% and 47.87% respectively. The metal ions biosorption was high at 4, 5-5.

Properties of Klebsiella Phage P13 and Associated Exopolysaccharide Depolymerase

The bacteriophage P13 that infects Klebsiella serotype K13 contains a heat-stable depolymerase capable of effective degradation of exopolysaccharide(EPS) produced by this microorganism. In this study, the titer of phage P13, initially 2.0 × 107 pfu mL-1, was found increasing 20 min after infection and reached 5.0 × 109 pfu mL-1 in 60 min. Accordingly, the enzyme activity of depolymerase approached the maximum 60 min after infection. Treatment at 70℃ for 30 min inactivated all the phage, but retained over 90% of the depolymerase activity. Addition of acetone into the crude phage lysate led to precipitation of the protein, with a marked increase in bacterial EPS degradation activity and a rapid drop in the titer of phage. After partial purification by acetone precipitation and ultrafiltration centrifugation, the enzyme was separated from the phage particles, showing two components with enzyme activity on Q-Sepharose Fast Flow. The soluble enzyme had an optimum degradation activity at 60℃ and pH 6.5. Transmission electron microscopy demonstrated that the phage P13 particles were spherical with a diameter of 50 nm and a short stumpy tail. It was a double-strand DNA virus consisting of a nucleic acid molecule of 45976 bp. This work provides an efficient purification operation including thermal treatment and ultrafiltration centrifugation, to dissociate depolymerase from phage particles. The characterization of phage P13 and associated EPS depolymerase is beneficial for further application of this enzyme.

Exopolysaccharides of Lactobacillus rhamnosus GG ameliorate Salmonella typhimurium-induced intestinal inflammation via the TLR4/NF-κB/MAPK pathway

Background Salmonella typhimurium(S.T),as an important foodborne bacterial pathogen,can cause diarrhea and gastroenteritis in humans and animals.Numerous studies have confirmed that exopolysaccharides(EPSs)have various biological functions,but the mechanism through which EPSs improve the immunity of animals against the invasion of pathogenic bacteria is unclear.Here,we explored the protective effect of EPSs of Lactobacillus rhamnosus GG(LGG)on the S.T-infected intestine.Methods Mice received adequate food and drinking water for one week before the start of the experiment.After 7 d of prefeeding,2×108 CFU/mL S.T solution and an equivalent volume of saline(control group)were given orally for 1 d.On the fourth day,the mice were treated with 0.5 mg/mL EPSs,1.0 mg/mL EPSs,2.0 mg/mL EPSs,or 2.0 mg/mL penicillin for 7 d.Finally,the body and relative organ weight,histological staining,and the levels of antioxidant enzyme activity and inflammatory cytokines were determined.Results The S.T-infected mice exhibited symptoms of decreased appetite,somnolence,diarrhea and flagging spirit.Treatment with EPSs and penicillin improved the weight loss of the mice,and the high dose of EPSs showed the best therapeutic effect.EPSs significantly ameliorated S.T-induced ileal injury in mice.High-dose EPSs were more effective than penicillin for alleviating ileal oxidative damage induced by S.T.The mRNA levels of inflammatory cytokines in the ileum of mice showed that the regulatory effects of EPSs on inflammatory cytokines were better than those of penicillin.EPSs could inhibit the expression and activation of key proteins of the TLR4/NF-κB/MAPK pathway and thereby suppress the level of S.T-induced ileal inflammation.Conclusions EPSs attenuate S.T-induced immune responses by inhibiting the expression of key proteins in the TLR4/NF-κB/MAPK signaling pathway.Moreover,EPSs could promote bacterial aggregation into clusters,which may be a potential strategy for reducing the bacterial invasion of intestinal epithelial cells.

Optimization of medium and cultivation conditions for enhanced exopolysaccharide yield by marine Cyanothece sp. 113

Cyanothece sp. 113 is a unicellular, aerobic, diazotrophic and photosynthetic marine cyanobacterium. The optimal medium for exopolysaccharide yield by the strain was 70.0 g/L of NaCl, and 0.9 g/L of MgSO4 based on the modified F/2 medium for cultivation of marine algae. The optimal cultivation condition for exopolysaccharide yield by this cyanobacterial strain was 29℃, aeration, and continuous illumination at 86.0 μE/M^2/S. Under the optimal conditions, over 18.4 g/L of exopolysaccharide was produced within 12 days. This was so far the highest exopolysaccharide yield produced with strains of Cyanothece sp. obtained.

Transcriptomic analysis reveals the effect of the exopolysaccharide of Psychrobacter sp.B-3 on gene expression in RAW264.7 macrophage cells

B-3 exopolysaccharide is extracted from the Antarctic psychrophilic bacterium Psychrobacter sp. B-3. We have previously shown that it activates macrophages and affects their immunoregulatory activities. To determine what genes are affected during this process, we detected the genes differentially expressed in cells of RAW264.7 macrophages treated with B-3 exopolysaccharide by transcriptomic analysis. B-3 exopolysaccharide treatment caused differential expression of 420 genes, of which 178 were up-regulated and 242 were down-regulated. These genes were shown to be involved in many aspects of cell function, mainly metabolism and immunity. Genes were enriched in multiple immune-related pathways, and the most significantly enriched genes were involved in antigen processing and presentation pathways. The pathway in which differentially expressed genes were the most significantly enriched was the metabolic pathway; specifically, the expression of many metabolic enzyme genes was altered by B-3 exopolysaccharide treatment. Additionally, the genes involved in metabolisms of amino acids, carbohydrates, lipids and nucleotides, varied to certain degrees. B-3 exopolysaccharide, therefore, appears to directly affect the immune function of RAW264.7 macrophages as an immunostimulant, or to indirectly change intracellular metabolism. This is the first study to determine the effect of an Antarctic psychrophilic bacterial exopolysaccharide on RAW264.7 macrophages. Our findings provide an important reference for research into the regulation of macrophage immune function by different polysaccharides.

Preparation and properties of bacteriophage-borne enzyme degrading bacterial exopolysaccharide

Bacteriophages infected different serotypes of Klebsiella were isolated from sewage. Among them, a heatstable polysaccharide depolymerase enzyme which could degrade bacterial exopolysaccharide effectively was prepared from the phage infecting Klebsiella K13. Treatment at 60℃ for 30 min could inactivate most of the K13 phage, with the titration decreasing from 6.4×10^8 PFU/mL to 1.6×10^6 PFU/mL. However, no obvious loss of phage enzyme activity was found after this treatment. The optimum hydrolytic temperature of phage enzyme was 60℃, with an activity 57 % higher than that at 30℃. The addition of phage enzyme could result in a rapid decrease of viscosity of exopolysaccharide （EPS） solution within minutes, indicating that K13 phage polysaccharide depolymerase acts as a kind of endo-glycanohydrolase. HPLC and reducing sugar analysis showed that the hydrolysis of EPS approached approximately the maxi-mum at 4h when the final concentration of phage was 6.0 x los PFU/mL. The results showed that K/eb-siella K13 phage depolymerase enzyme could be used as a good tool for the preparation of EPS oligosac- charide.

Exopolysaccharides Produced from <i>Lactobacillus delbrueckii</i>subsp. <i>bulgaricus</i>

Lactobacillus delbrueckii subsp. bulgaricus, which has been widely used as a fermented milk starter, is a type of probiotic, and certain strains are able to produce exopolysaccharide (EPS). EPS produced from L. bulgaricus contributes to the physical and biological function of dairy products by regulating immune response, and this tendency seems to place EPS with acidic groups. To date, six types of chemical structure have been determined and are basically composed from glucose (Glc), galactose (Gal), and rhamnose (Rha). Eps clusters on chromosome DNA control the EPS synthesis and are transcribed as one mRNA 14 genes with 18kb on L. bulgaricus Lfi5. Furthermore, L. bulgaricus is able to utilize lactose (Lac) as carbohydrate source, repeating units of EPS are synthesized from Glc 6-phosphate, generated by an Embden-Meyerhof (EM) pathway in cellular carbohydrate assimilation. This review discusses EPS produced from L. bulgaricus.

Antioxidant Activities of Exopolysaccharides Produced by Lactic Acid Bacteria Isolated from Commercial Yoghurt Samples

An antioxidant is a substance that inhibits the oxidation of other molecules caused by free radicals. The inbuilt antioxidant systems possessed by living organisms are generally not enough to prevent them from oxidative damages and the uses of synthetic antioxidants also have some harmful effects. This study was aimed at evaluating the antioxidant activities of exopolysaccharides produced by lactic acid bacteria isolated from yoghurt. Lactic acid bacteria (LAB) were isolated from six different brands of commercially available yoghurt using deMan Rogosa Sharpe (MRS) agar. The LAB isolates were identified based on morphological and biochemical analyses and were screened for exopolysaccharide (EPS) production. The LAB isolates screened positive were used for EPS production in a liquid medium and the EPS produced were purified and quantified using standard methods. Antioxidant activities of the EPS were evaluated by determining the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, ferric ion reducing power, and total phenolic contents. Data obtained were analysed using Analysis of Variance. Total lactic acid bacterial count obtained from the yoghurt samples ranged from 0 - 3.9 × 104 CFU/mL with sample A (Fan Yoghurt) having the highest LAB count (3.9 × 104 CFU/mL). The isolated LAB and their incidence rate were Lactobacillus plantarum (25.49%), L. delbrueckii (19.61%), L. fermentum (15.69%), L. acidophilus (13.73%), Leuconostoc mesenteroides (11.76%), Lactococcus lactis (7.84%), and Lactobacillus casei (5.88%). Fifty-one out of the 64 LAB isolates were screened positive for EPS production and only six were able to produce substantial quantity of EPS ranging from 127.4 - 208.5 mg/L. The exopolysaccharides produced by L. fermentum had the highest DPPH radical scavenging activity (62.90%) while that of L. plantarum had the lowest (23.10%) at a concentration of 1000 μg/mL. Also, the EPS produced by L. fermentum recorded the highest ferric ion reducing power (12.89 mg AAE/mL) at 1000 μg/mL while that of L. plantarum had the lowest (5.62 mg AAE/mL). At 1000 μg/mL, the total phenolic contents of the EPS samples ranged from 1.41 - 1.58 mg GAE/mL, and the EPS produced by L. fermentum had the highest (1.58 mg GAE/mL) while those produced by L. paracasei had the lowest (1.41 mg GAE/mL). This study revealed that the exopolysaccharides produced by the LAB isolates showed high antioxidant activities with respect to their DPPH free radical scavenging activity, ferric ion reducing power and total phenolic contents.

Activation of macrophages by an exopolysaccharide isolated from Antarctic P sychrobacter sp.B-3

An exopolysaccharide(EPS) was isolated and purified from an Antarctic psychrophilic bacterium B-3,identified as Psychrobacter sp.,and the activation of RAW264.7 cells by B-3 EPS was investigated.The results show that B-3 EPS,over a certain concentration range,promoted cell viability,nitric oxide production,tumor necrosis factor(TNF)a secretion,and phagocytic ability.Furthermore,TAK-242,an inhibitor of the toll-like receptor 4(TLR4) significantly reduced nitric oxide production by these cells after stimulation with B-3 EPS.Moreover,B-3 EPS induced p65 phosphorylation and IκBα degradation in these cells.In conclusion,B-3 EPS might have activated RAW264.7 cells by combining with TLR4 on cell surface and triggering activation of NF-κB signaling pathways,implying that this EPS could activate macrophages and regulate initial immune response.

Exopolysaccharide Production by Four Cyanobacterial Isolates and Preliminary Identification of These Isolates

Four marine cyanobacterial isolates, named 104, 109, 113 and 115, from marine water off China’s coast can release a large amount of exopolysaccharide (EPS) to medium.The effects of different components in medium on EPS production by the four isolates were investigated. Under the optimal condition, the EPS released by isolates 104, 109, 113 and 115 reached 7.48 g L-1, 8.33 g L-1, 18.26 g L-1, and 6.78 g L-1 within 14 d, respectively. Based on the conventional identification methods for cyanobacteria, these isolates were assigned to genus Cyanothece.

Structural Characterization of an Exopolysaccharide from the Myxobacterium Sorangium Cellulosum NUST06

The chemical structure of an exopolysaccharide (EPS) produced by Sorangium cellulosum NUST06 was investigated. The EPS was found to have molecular masses greater than 2?106 Da, and consisted of D-glucose, D-mannose and D-glucuronic acid in a ratio of 5:4:1. The analytical results of methylation and nuclear magnetic resonance spectra suggested the EPS consist of following tetrasaccharide repeating units: 4) -b-D-Glup (1 4)- b-D-Glup (1 4) -b-D-Glup (1 a-D-GluAp (1 6) a-D-manp (1 6)-a-D-manp (1 6) (6) (4) (4) ↑ ↑ ↑ Me Ac Ac

A Suitable Culture Medium for the Production of Exopolysaccharide by Streptococcus salivarius subsp.thermophilus

A new culture medium for the synthesis of exopolysaccharide by streptococcus salivarius subsp.thermophilus was formulated,based on MRS,M17,ATP,SL,Elliker,SDM usually used in lactic acid bacterium.The suitability of a semi defined culture media (named LCX) was obtained.The formula of culture medium was optimized as:peptone,1.50%,typtone,1.00%;K 2HPO 4,0.20%;MgSO 4.7H 2O,0.02%;MnSO 4.4H 2O,50 mg·L -1 ;Tween 80,1ml·L -1 ;Sodium acetate,0.50%;Sodium glycerate,1.90%.

Influence of Different Sterilization Conditions on the Growth and Exopolysaccharide of <i>Streptococcus thermophilus</i>and Co-Cultivation with <i>Lactobacillus delbrueckii</i>subsp. <i>bulgaricus</i>OLL1073R-1

The sterilization conditions between experiment and dairy industrial level are different and concern about influence on product’s qualities. In this study, the fermentation properties of Streptococcus thermophilus, alone and in combination with Lactobacillus delbrueckii subsp. bulgaricus, were evaluated in skim milk that had been subjected to distinct sterilization conditions. Growth, organic acid generation, and EPS production were determined using pasteurized or autoclaved milk. When S. thermophilus was cultivated in pasteurized skim milk, the growth was strain-dependent. On the other hand, growth of S. thermophilus was accelerated in autoclaved milk. Exocellular polysaccharide (EPS) production by L. bulgaricus was not affected by the combination of S. thermophilus strains. Thus, we observed that yogurt fermented by L. bulgaricus was minimally affected by the combination of S. thermophilus strains;growth of L. bulgaricus was maintained under the constant environment. These results should facilitate the development of fermented milk produced from L. bulgaricus in the dairy industry.

Studies on the optimal culture conditions of <i>Aureobasidium pullulans</i>to produce exopolysaccharides

In the current study, in order to change the permeability of cell membrane and solve the problem of linked group of fungi mycelium, the method of adjusting osmotic pressure of medium and adding tween-80 was established. The utilized strain with relatively high exopolysaccharide (EPS) yield and low pigment level was obtained after the rejuvenation and sifting of long-preserved Aureobasidium pullulans strain. The optimal proportion of substrate was determined by means of orthogonal test. The transformation ratio of EPS was increased by 10% - 20% and the pigment content was greatly reduced. The fermenting liquor is between creamy white and pale yellow, and the white primary product can be gained without decolourization step. Furthermore, to magnify to 5 L bioreactor can get the similar result.

Removal of Silver from Aqueous Solution by <i>Azotobacter chroococcum</i>XU1 Biomass and Exopolysaccharide

Biosorption of silver ions onto A. chroococcum XU1 biomass and exopolysaccharide (EPS) was investigated. It was found that the overall biosorption process was the best when cell biomass and EPS were used together. Metal adsorption experiments showed that the final precipitate obtained by cell biomass and EPS, contained 7.85% of Ag. Based on the results presented in this study, the biomass and EPS of A. chroococcum XU1 indicated the possibility of application of them as biosorbent for removal of silver from waste waters.