• Title/Summary/Keyword: Exopolysaccharides (EPS)

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Effect of Ammonium Phosphate on Mycelial Growth and Exopolysaccharides Production of Ganoderma lucidum in an Air-Lift Fermenter

  • Lee, Kyu-Min;Lee, Shin-Young;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.726-731
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    • 1999
  • It was discovered that ammonium phosphate in the medium played an important role in both growing mycelium and producing exopolysaccharides (EPS) from G. lucidum. In lower concentration levels of ammonium phosphate (0-3 g/l), an improved mycelial growth was observed by maintaining more filamentous morphology than in high concentrations (5-11 g/l). In addition, it was confirmed by comparing the factual dimension and frequency of the area regarding the mycelial pellets. This must be attributed to limitations of nutrient transfer by maintaining filamentous mycelium during the cultivation in a low ammonium phosphate containing medium. On the other hand, the best EPS production was observed in medium with the absence or low concentration of ammonium phosphate. The shear stress of the culture broth was greatly affected by the shear rate, as compared with that of the culture broth with high ammonium phosphate concentration. The rheological characteristics of the fermentation broth and filtrate worked well according to the Herschel-Bulkley model. It was also found that the morphological changes of the mycelium resulting from the ammonium phosphate concentration directly affected the rheological characteristics of the system and resulted in reversely affecting the EPS production levels. Based on these results, it can be concluded that delicate regulation of the ammonium phosphate concentration in the culture media should be provided in order to obtain optimal mycelial growth and/or EPS production.

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Inhibitory Effect on Rotavirus by Exopolysaccharides Extracted from Kefir (Kefir에서 추출한 Exopolysaccharide의 Rotavirus의 저해효과)

  • Song, Jin-Ook;Kim, Tae-Jin;Kim, Yong-Hui
    • Food Science of Animal Resources
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    • v.27 no.4
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    • pp.538-542
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    • 2007
  • This study was carried out to investigate the toxicity of exopolysaccharides (EPS) from kefir toward MA104 cells and evaluate the inhibitory effects of kefir EPS on rotavirus infection. The results obtained are summarized as follows: Lactic acid bacteria (Lactobacillus fermentum, L. acidophilus, L. brevis) and yeasts (Candida kefyr, Cryptococcus albidus, Pichia ohmeri) were isolated and identified from kefir grain and culture. At 1% EPS, the inhibitory effects of EPS on the infection of MA-104 cells using the MTT assay were $72.52{\pm}6.48%$ for human rotavirus (KU), $36.06{\pm}7.63%$ for bovine rotavirus (NCDV), and $81.66{\pm}1.11%$ for porcine rotavirus (OSU). At 1/128% EPS, the effects were $24.98{\pm}4.58%$ for human rotavirus (KU), $4.71{\pm}6.16%$ for bovine rotavirus (NCDV), and $4.05{\pm}14.90%$ forporcine rotavirus (OSU). EPS isolated from kefir have inhibitory effects on rotaviruses of various serotypes and rotaviruses from different animals.

Function of Cell-Bound and Released Exopolysaccharides Produced by Lactobacillus rhamnosus ATCC 9595

  • Kim Ji-Uk;Kim Young-Hoon;Han Kyoung-Sik;Oh Se-Jong;Whang Kwang-Youn;Kim Jai-Neung;Kim Sae-Hun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.939-945
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    • 2006
  • The physiological characteristics and function of the exopolysaccharide (EPS) produced by Lactobacillus rhamnosus ATCC 9595 were determined. The total quantity of EPS was rapidly increased to 496$\pm$20 mg/l during the exponential phase, and then maintained steadily during the stationary phase. During the exponential phase (18 h), the total EPS consisted of 61% cell-bound EPS (cb-EPS) and 39% released EPS (r-EPS), whereas the relative proportion of EPS during the stationary phase (48 h) was convered to 23% cb-EPS and 77% r-EPS. On gel permeation chromatography, cb-EPS was fractionated as a single peak of 8.6$\times10^6$ Da, whereas r-EPS was fractionated as two peaks with average molecular weights of 4.3$\times$10$^4$ and 8.6$\times10^6$ Da. Interestingly, both EPS species exhibited anticancer properties and cholera toxin-binding activities. Our results suggest that the EPS generated by L. rhamnosus ATCC 9595 might be suitable for use as a functional food or food supplement.

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.

Production of Exopolysaccharides by Lactococcus lactis subsp. cremoris MG1363 Expressing the eps Gene Clusters from Two Strains of Lactobacillus rhamnosus

  • Kang, Hye-Ji;LaPointe, Gisele
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.91-101
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    • 2018
  • The aim of this study was to transfer the 18.5 kb gene clusters coding for 17 genes from Lactobacillus rhamnosus to Lactococcus lactis subsp. cremoris MG1363 in order to determine the effect of host on exopolysaccharide (EPS) production and to provide a model for studying the phosphorylation of proteins which are proposed to be involved in EPS polymerization. Lactobacillus rhamnosus RW-9595M and ATCC 9595 have 99% identical operons coding for EPS biosynthesis, produced different amounts of EPS (543 vs 108 mg/l). L. lactis subsp. cremoris MG1363 transformed with the operons from RW-9595M and ATCC 9595 respectively, produced 326 and 302 mg/l EPS in M17 containing 0.5% glucose. The tyrosine protein kinase transmembrane modulator (Wzd) was proposed to participate in regulating chain elongation of EPS polymers by interacting with the tyrosine protein kinase Wze. While Wzd was found in phosphorylated form in the presence of the phosphorylated kinase (Wze), no phosphorylated proteins were detected when all nine tyrosines of Wzd were mutated to phenylalanine. Lactococcus lactis subsp. cremoris could produce higher amounts of EPS than other EPS-producing lactococci when expressing genes from L. rhamnosus. Phosphorylated Wzd was essential for the phosphorylation of Wze when expressed in vivo.

Structure and Foaming Properties of Viscous Exopolysaccharides from a Wild Grape-Associated Basidiomycetous Yeast Papiliotrema flavescens Formerly Known as Cryptococcus flavescens

  • Oluwa, Salomon Woye
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1739-1749
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    • 2020
  • Exopolysaccharide produced by the yeast Papiliotrema flavescens, isolated from wine grape berries of Champagne vineyard, was investigated for both chemical and functional characterization. SECMALLS and colorimetric assay analyses showed that the EPS is a high MW heteropolymer (2.37 × 106 g/mol) majorily consisting of mannose, glucose, xylose and glucuronic acid as monosaccharide constituents, with two substituents (sulphate and phosphate groups), and a minor protein moiety. Structural enchainment of these carbohydrates based on methylation, GC-MS and NMR analyses revealed a linear main backbone built up of α-(1 → 3)-D-mannopyranosyl residues on which are branched side chains consisting of a single β-D-glucopyranosyluronic acid residue and β-(1 → 2)-xylopyranoses (2-5 residues). Suggestion of some xylopyranose side chains containing a mannose residue at the nonreducing terminal end was also proposed. This is first report on EPSs from the grape P. flavescens yeast with such structural characteristics. Furthermore, investigations for valuating the application performance of these EPS in relation with their structural features were carried out in 8% alcohol experiment solutions. Very exceptional viscosifying and foaming properties were reported by comparison with commercial biopolymers such as Arabic, gellan and xanthan gums. The intrinsic properties of the natural biopolymer from this wild grape-associated P. flavescens yeast make it a potential candidate for use in various biotechnology applications.

Isolation and Characterization of Mucous Exopolysaccharide (EPS) Produced by Vibrio furnissii Strain VB0S3

  • Bramhachari P.V.;Kishor P.B. Kavi;Ramadevi R.;Kumar Ranadheer;Rao, B. Rama;Dubey Santosh Kumar
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.44-51
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    • 2007
  • Marine bacterial strains were isolated trom coastal regions of Goa and screened for the strains that produce the highest amount of mucous expolysaccharide (EPS). Our screening resulted in the identification of the strain Vibrio furnissii VB0S3 (hereafter called VB0S3), as it produced the highest EPS in batch cultures during the late logarithmic growth phase. The isolate was identified as VB0S3 based on morphological and biochemical properties. Growth and EPS production were studied in mineral salts medium supplemented with NaCl (1.5%) and glucose (0.2%). The exopolymer was recovered from the culture supernatant by using three volumes of cold ethanol precipitation and dialysis procedure. Chemical analyses of EPS revealed that it is primarily composed of neutral sugars, uronic acids, and proteins. Fourier-transform infrared (FT-IR) spectroscopy revealed the presence of carboxyl, hydroxyl, and amide groups, which correspond to a typical heteropolymeric polysaccharide, and the EPS also possessed good emulsification activity. The gas chromatographic analysis of an alditol-acetate derivatized sample of EPS revealed that it was mainly composed of galactose and glucose. Minor components found were mannose, rhamnose, fucose, ribose, arabinose, and xylose. EPS was readily isolated from culture supernatants, which suggests that the EPS was a slime-like exopolysaccharide. This is the first report of exopolysaccharide characterization that describes the isolation and characterization of an EPS expressed by Vibrio surnissii strain VB0S3. The results of the study contribute significantly and go a long way towards an understanding of the correlation between growth and EPS production, chemical composition, and industrial applications of the exopolysaccharide in environmental biotechnology and bioremediation.

Kinetic Modeling of Submerged Culture of A. blazei with Mixed Carbon Sources of Glucose and Dextrin

  • Na Jeong-Geol;Kim Hyun-Han;Chang Yong-Keun;Lee Sang-Jong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1331-1337
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    • 2006
  • A mathematical model has been proposed for the batch culture of Agaricus blazei with mixed carbon sources of glucose and dextrin. In the proposed model, the metabolism of A. blazei was divided into three parts: cell growth, exopolysaccharides (EPS) production, and another EPS production pathway activated by dextrin hydrolysis. Each pathway was described mathematically and incorporated into the mechanistic model structure. Batch cultures were carried out with six different carbon source compositions. Although parameters were estimated by using the experimental data from the two extreme cases such as glucose only and dextrin only, the model represented well the profiles of glucose, cell mass, and EPS concentrations for all the six different carbon source mixtures, showing a good interpolation capability. Of note, the lag in EPS production could be quite precisely predicted by assuming that the glucose-to-cell mass ratio was the governing factor for EPS production.

Supercooling Pretreatment Improves the Shelf-Life of Freeze-Dried Leuconostoc mesenteroides WiKim32

  • Seul-Gi Jeong;In Seong Choi;Ho Myeong Kim;Ji Yoon Chang;Hae Woong Park
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1599-1604
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    • 2022
  • Storage stability of freeze-dried lactic acid bacteria is a critical factor for their cost-effectiveness. Long-term storage of lactic acid bacteria enables microbial industry to reduce distribution costs. Herein, we investigated the effect of cold adaptation under supercooling conditions at -5℃ on the viability of Leuconostoc mesenteroides WiKim32 during the freeze-drying process and subsequent storage. Cold adaptation increased the thickness of exopolysaccharides (EPS) and improved the viability of freeze-dried Leu. mesenteroides WiKim32. Compared to non-adapted cells, cold-adapted cells showed a 35.4% increase in EPS thickness under supercooling conditions. The viability of EPS-hydrolyzed cells was lower than that of untreated cells, implying that EPS plays a role in protection during the freeze-drying process. Cold adaptation increased the storage stability of freeze-dried Leu. mesenteroides WiKim32. Fifty-six days after storage, the highest viability (71.3%) was achieved with cold adaptation at -5℃. When EPS-containing broth was added prior to the freeze-drying process, the viability further increased to 82.7%. These results imply that cold adaptation by supercooling pretreatment would be a good strategy for the long-term storage of Leu. mesenteroides WiKim32.

Overexpression of Capsular Polysaccharide Biosynthesis Protein in Lactobacillus plantarum P1 to Enhance Capsular Polysaccharide Production for Di-n-butyl Phthalate Adsorption

  • Liu, Wei-Bing;Lin, Zhi-Wei;Zhou, Ying;Ye, Bang-Ce
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1545-1551
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    • 2021
  • Exopolysaccharides (EPSs) such as capsular polysaccharide (CPS) are important bioactive carbohydrate compounds and are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPS-producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs having many different structures and no health risks. However, EPS production by LAB does not meet the needs of large-scale application on an industrial scale. Here, the capA gene (encoding CPS biosynthesis protein) was overexpressed in Lactobacillus plantarum P1 to improve the production of EPSs and further enhance the DBP adsorption capability. Compared with P1, the CPS production in capA overexpressed strain was increased by 11.3 mg/l, and the EPS thickness was increased from 0.0786 ± 0.0224 ㎛ in P1 to 0.1160 ± 0.0480 ㎛ in P1-capA. These increases caused the DBP adsorption ratio of P1-capA to be doubled. Overall, the findings in this study provide a safe method for the adsorption and removal of DBP.