• Title/Summary/Keyword: exopolysaccharide (EPS)

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Chemical Modification of Botryosphaeran: Structural Characterization and Anticoagulant Activity of a Water-Soluble Sulfonated ($1{\rightarrow}3$)($1{\rightarrow}6$)-${\beta}$-D-Glucan

  • Brandi, Jamile;Oliveira, Eder C.;Monteiro, Nilson K.;Vasconcelos, Ana Flora D.;Dekker, Robert F.H.;Barbosa, Aneli M.;Silveira, Joana L.M.;Mourao, Paulo A.S.;Silva, Maria De Lourdes Corradi Da
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1036-1042
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    • 2011
  • The exopolysaccharide botryosphaeran ($EPS_{GLC}$; a ($1{\rightarrow}3$)($1{\rightarrow}6$)-${\beta}$-D-glucan from Botryosphaeria rhodina MAMB-05) was sulfonated to produce a water-soluble fraction ($EPS_{GLC}$-S) using pyridine and chlorosulfonic acid in formamid. This procedure was then repeated twice to produce another fraction ($EPS_{GLC}$-RS) with a higher degree of substitution (DS, 1.64). The purity of each botryosphaeran sample (unsulfonated and sulfonated) was assessed by gel filtration chromatography (Sepharose CL-4B), where each polysaccharide was eluted as a single symmetrical peak. The structures of the sulfonated and re-sulfonated botryosphaerans were investigated using ultraviolet-visible (UV-Vis), Fourier-transform infrared (FT-IR), and $^{13}C$ nuclear magnetic resonance ($^{13}C$ NMR) spectroscopies. $EPS_{GLC}$ and $EPS_{GLC}$-RS were also assayed for anticoagulation activity, and $EPS_{GLC}$-RS was identified as an anticoagulant.

Exopolysaccharide-Overproducing Lactobacillus paracasei KB28 Induces Cytokines in Mouse Peritoneal Macrophages via Modulation of NF-${\kappa}B$ and MAPKs

  • Kang, Hee;Choi, Hye-Sun;Kim, Ji-Eun;Han, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1174-1178
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    • 2011
  • Exopolysaccharides (EPSs) are microbial polysaccharides that are released outside of the bacterial cell wall. There have been few studies on EPS-producing lactic acid bacteria that can enhance macrophage activity and the underlying signaling mechanism for cytokine expression. In the current study, EPS-overproducing Lactobacillus (L.) paracasei KB28 was isolated from kimchi and cultivated in conditioned media containing glucose, sucrose, and lactose. The whole bacterial cells were obtained with their EPS being attached, and the cytokine-inducing activities of these cells were investigated. Gas chromatography analysis showed the presence of glucose, galactose, mannose, xylose, arabinose, and rhamnose in EPS composition. EPS-producing L. paracasei KB28 induced the expression of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12 in mouse macrophages. This strain also caused the degradation of $I{\kappa}B{\alpha}$ and phosphorylation of the major MAPKs: Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK)1/2. The use of pharmacological inhibitors showed that different signaling pathways were involved in the induction of TNF-${\alpha}$, IL-6 and IL-12 by L. paracasei KB28. Our results provide information for a better understanding of the molecular mechanisms of the immunomodulatory effect of food-derived EPS-producing lactic acid bacteria.

Aglycone Isoflavones and Exopolysaccharides Produced by Lactobacillus acidophilus in Fermented Soybean Paste

  • Kim, Jin-Sun;Lee, Je-Hyuk;Surh, Jeonghee;Kang, Soon Ah;Jang, Ki-Hyo
    • Preventive Nutrition and Food Science
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    • v.21 no.2
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    • pp.117-123
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    • 2016
  • Bioconversion of aglycone-formed isoflavones from glycoside-formed isoflavones by commercial lactic acid bacteria in fermented soybean paste was evaluated. Enterococcus faecium KCTC 13410 showed the most resistant capacity and Lactobacillus acidophilus KCTC 3925 had a sensitive susceptibility at a high NaCl concentration (13.2%) in fermented soybean paste. Among the 5 strains tested, Lac. acidophilus KCTC 3925 showed the highest relative ratio of aglycone-formed isoflavones to total isoflavones in fermented soybean paste. Production of exopolysaccarides (EPS) by lactic acid bacteria was compared using de Man, Rogosa, and Sharpe medium containing 1% sucrose at $37^{\circ}C$ for 48 h. Among the 5 lactic acid bacteria, Lac. acidophilus KCTC 3925 and Lactobacillus rhamnosus KCTC 3929 were investigated to produce EPS. Based on the results concerning growing susceptibility and conversion of aglycone-formed isoflavones/EPS production, it is anticipated that Lac. acidophilus KCTC 3925 may be used for preparation of Cheonggukjang, which contains relative low NaCl content.

Heat-Killed Enterococcus faecium KU22001 Having Effective Anti-Cancer Effects on HeLa Cell Lines at a Lower Temperature

  • Jun-Su Ha;Na-Kyoung Lee;Hyun-Dong Paik
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.902-910
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    • 2024
  • The anti-cancer effects of heat-killed Enterococcus faecium KU22001 (KU22001), KU22002, and KU22005 isolated from human infant feces were investigated. The anti-proliferative activity of these strains against various cancer cell lines was evaluated using the MTT assay. To determine the production of exopolysaccharides (EPS) with potential anti-cancer effect, ethanol precipitation and phenol-sulfuric acid method was used with the cell free supernatant of strains grown at 25℃ or 37℃. The EPS yield of E. faecium strains was higher at 25℃ than at 37℃. Among these E. faecium strains, KU22001 grown at 25℃ was associated with the highest bax/bcl-2 ratio, effective apoptosis rate, cell cycle arrest in the G0/G1 phase, and condensation of the nucleus in the cervical cancer HeLa cell line. In conclusion, these results suggest that KU22001 can be beneficial owing to the anti-cancer effects and production of functional materials, such as EPS.

Isolation and Characterization of Exopolysaccharide-Producing Bacteria from Korean Fermented Vegetables (전통 침채류 유래 다당 생산균의 분리 특성)

  • Kwon, Tae-Yeon;Shim, Sang-Min;Heo, Min-Young;An, Doo-Hyun;Shin, Kwang-Soon;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.191-195
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    • 2007
  • Four bacteria producing viscous exopolysaccharides (EPSs) were isolated from Korean fermented vegetables (Cucumber kimchi, Young radish kimchi, Green onion kimchi) using a selection medium intended for isolating bacteria with tannin-degrading activity. They were identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Enterobacter cowan ii, Escherichia senegalensis, Enterobacter asburiae, and Enterobacter ludwigii. Strain CK31, the most efficient EPS-producer, produced a heteropolysaccharide with an approximate molecular weight of 420 kDa. The neutral sugar fraction of the EPS was composed of rhamnose, fucose, arabinose, mannose, galactose, and glucose.

Influence of Agitation Intensity and Aeration Rate on Production of Antioxidative Exopolysaccharides from Submerged Mycelial Culture of Ganoderma resinaceum

  • Kim Hyun-Mi;Kim Sang-Woo;Hwang Hye-Jin;Park Moon-Ki;Mahmoud Yehia A.-G.;Choi Jang-Won;Yun Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1240-1247
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    • 2006
  • The present study investigated the influence of the aeration rate and agitation intensity on the production of the mycelial biomass and antioxidative exopolysaccharide (EPS) in Ganoderma resinaceum. In submerged cultures with varying agitation speeds and aeration rates in a stirred-tank reactor, the maximum mycelial biomass and maximum EPS concentration were achieved at 50 rpm and 300 rpm, respectively. Under varying aeration rates, the highest amount of mycelial biomass (18.1 g/l) was accumulated at the lowest aeration rate (0.5 vvm) and the maximum EPS production (3.0 g/l) obtained at 1.0 vvm. A compositional analysis revealed that the five different EPSs were protein-bound heteropolysaccharides, consisting of 87.17-89.22% carbohydrates and 10.78-12.83% proteins. The culture conditions had a striking affect on the carbohydrate composition of the EPS, resulting in different antioxidative activities. All the EPSs showed strong scavenging activities against superoxide and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals, whereas no clear trend in antioxidative activity was observed against hydroxyl radicals and lipid peroxides. Although the precise reason for this difference is still unclear, the high glucose moiety of EPS is probably linked to its broad spectrum of antioxidative activity.

Physicochemical Properties of the Exopolysaccharides Produced by Marine Bacterium Zoogloea sp. KCCM10036

  • Lim, Dong-Jung;Kim, Jong-Deog;Kim, Min-Yong;Yoo, Sang-Ho;Kong, Jai-Yul
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.979-984
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    • 2007
  • The physicochemical properties of the exopolysaccharide (EPS) produced by marine bacterium Zoogloea sp. KCCM10036 were investigated. Two types of isolated EPSs were shown to have average relative molecular masses $(M_r)\;of\;4.07{\times}10^6$ of CBP (cell-bound polysaccharide) and $3.43{\times}10^6$ of WSP (water-soluble polysaccharide), respectively. When the CBP was utilized as an emulsifier, it stabilized the emulsifier, for up to 148 h. Compared with other commercially available hydrocolloids such as xanthan gum, the Tween series, and Triton, the CBP showed much better emulsifying capability on a water-in-oil system. Phase separation occurred in the Tween series after 24 h, whereas the emulsion was better stabilized by the CBP. The CBP thus has potential as an emulsifying agent in commercial emulsions. The flocculating activity was also greatest at 0.01% (w/v) and decreased at higher concentrations than the optimized concentration of the WSP and CBP. The results also showed that both types of expolysaccharides from Zoogloea sp. had excellent flocculating activity.

Metabolic Flux Analysis of Beijerinckia indica for PS-7 Production

  • Wu Jian-Rong;Son Jeong Hwa;Seo Hyo-Jin;Kim Ki-Hong;Nam Yoon-Kwon;Lee Jin-Woo;Kim Sung-Koo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.91-98
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    • 2005
  • In order to investigate central metabolic changes in Beijerinckia indica, cells were grown on different carbon sources and intracellular flux distributions were studied under varying concentrations of nitrogen. Metabolic fluxes were estimated by combining material balances with extracellular substrate uptake rate, biomass formation rate, and exopolysaccharide (EPS) accumulation rate. Thirty-one metabolic reactions and 30 intracellular metabolites were considered for the flux analysis. The results revealed that most of the carbon source was directed into the Entner-Doudoroff pathway, followed by the recycling of triose-3-phosphate back to Hexose­6-phosphate. The pentose phosphate pathway was operated at a minimal level to supply the precursors for biomass formation. The different metabolic behaviors under varying nitrogen concentrations were observed with flux analysis.

Potential Probiotic Properties of Exopolysaccharide Producing Lactic Acid Bacteria Isolated from Fermented Soybean Product (장류유래 Exopolysaccharide 생성 유산균의 잠재적 Probiotic 특성)

  • Ahn, Yu-Jin;Choi, Hye-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.749-755
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    • 2014
  • Exopolysaccharides (EPSs) have been widely used in the food industry as viscofying, stabilizing, and emulsifying agents as well as in the pharmaceutical industry for their immunomodulatory, anti-tumor, and anti-inflammatory effects. A total of 458 lactic acid bacteria (LAB) strains isolated from several kinds of soybean pastes were screened for the production of homo-EPS (HoPS). LAB isolates were primarily screened using thin layer chromatography (TLC) and further screened polymerase chain reaction (PCR) targeting genes involved in HoPS production. Six LAB isolates producing high amounts of HoPS were identified by TLC. Among these isolates, glucansucrase gene was amplified in two strains (JSA57, JSB22), whereas the fructansucrase gene was detected in three strains (JSA57, JSB22, JSB66). After isolating the strains, their morphological characteristics and 16S rDNA sequences were determined. Six species were identified as L. alimentarius HSB15, L. plantarum JSA22, L. pentosus JSA57, L. brevis JSB22, L. alimentarius JSB66, and L. parabrevis JSB89. To evaluate the potential probiotic properties of these LAB, their survival rates against a simulated intestinal environment were determined. After 2 hr of incubation in artificial gastric juice, survival rates of JSA57, JSB90, JSB22, and JSB66 were all greater than 50%. After 2 hr of incubation in bile juice, viable cell count of JSB22 was similar with initial vial cell counts. Growth of the six LAB was screened in arabino-oligosaccharide (AOS)-containing MRS broth. Results showed that growth of the isolates selectively increased after culture in AOS-containing media. Strain JSB22 (6 hr), JSB66 (6 hr), HSB15 (20 hr), and JSA22 (29 hr) showed maximum growth rate. Especially, JSB22 showed the highest growth rate. These results suggest that EPS-producing LAB isolated from Deonjang could be applied as synbiotics.

Cloning and Sequencing of a Gene Involved in the Biosynthesis of Exopolysaccharide in Zoogloea Ramigera 115SLR (Zoogloea Ramigera 115SLR로부터 다당류 생합성에 관여하는 유전자의 분리 및 염기서열 결정)

  • Sam-Pin Lee;Min Yoo
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.1-9
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    • 2000
  • To identity the genes responsible for the biosynthesis of exopolysaccharide, recombinant plasmids pUEX10 and pLEX10 were constructed from plasmid pLEX3 which was isolated from the recombinant cosmid library of Zoogloea ramigera 115. The complete nucleotide sequence of the 1.7 kb genomic DNA insert in plasmid pUEX10 was determined. Its analysis identified two open reading frames (ORF3 & ORF4) which could encode two proteins. The amino acid sequence derived from ORF3 showed the homology with gumC protein in Xanthomonas campestris as well as exoP protein in Rhizobium melizoti. The partial amino acid sequence of ORF4 showed the homology with polysaccharide export protein in Thermotoga maritima. Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 showed the similar pattern for EPS production. Yield of exopolysaccharides produced by Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 was 0.26% (w/v) and 0.16% (w/v), respectively.

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