• Title/Summary/Keyword: exopolysaccharide EPS-1

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Inhibition of Quorum Sensing Regulated Virulence Factors and Biofilm Formation by Eucalyptus globulus against Multidrug-Resistant Pseudomonas aeruginosa

  • Sagar, Pankaj Kumar;Sharma, Poonam;Singh, Rambir
    • Journal of Pharmacopuncture
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    • v.25 no.1
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    • pp.37-45
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    • 2022
  • Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.

Optimal Conditions of Mycelial Growth and Exopolysaccharide Production in Submerged Culture of Phellinus baumii (Phellinus baumii으로부터 세포외 다당체 생산의 최적화)

  • Hwang, Hye-Jin;Kim, Sang-Woo;Yun, Jong-Won;Park, Jang-Won
    • Journal of Life Science
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    • v.14 no.1
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    • pp.51-56
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    • 2004
  • The polysaccharide isolated from Phellinus species has been known as a folk remedy, including antitumor and immune-stimulating activities. However, there are lacks of knowledge about mycelial growth and exopolysaccharide (EH) production in its submerged culture. We investigated the optimal conditions on mycelial growth and EPS production in Phellinus baumii. The optimal temperature and initial pH for mycelial growth and EPS production in shake flask culture of P. baumii were proved to be 3$0^{\circ}C$ and pH 5.0, respectively. In case of carbon source, cellobiose and maltose were highly efficient for mycelial growth and fructose and mannitol were also relatively favorable for EPS production. Yeast extract was the most suitable nitrogen source for mycelial growth and EPS production. The composition of optimal culture medium was determined to be fructose 20 g/L, yeast extract 20 g/L, and $CaCl_2$ 0.55 g/L, respectively. Under the optimal culture condition, the maximum mycelial biomass and EPS achieved in a 5-L stirred-tank fermenter were 17.43 g/L and 3.6 g/L, respectively. It was found that the EPS was a glycoprotein onsisted of mainly arginine (14.1%) and glycine (12.0 %) in protein moiety and mainly mannose (48.7%) and arabinose (38.4%) in carbohydrate moiety.

Anti-diabetic Effect of the Exopolysaccharides (EPS) Produced from Cordyceps sinensis on ob/ob Mice (제 2형 당뇨쥐에서 동충하초로부터 생산된 세포외 다당류의 항당뇨 효과)

  • Choi, Jang-Won
    • KSBB Journal
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    • v.26 no.1
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    • pp.33-40
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    • 2011
  • Anti-diabetic effect of the exopolysaccharides (EPS) produced from submerged mycelial culture of Cordyceps sinensis (Cs) was studiedin a type II diabetic animal model (C57BL/6J ob/ob). This study was designed to determine whether Cs-EPS improves clinical symptoms of type II diabetes in ob/ob mice. After Cs-EPS treatment at doses of 200 mg/kg body weight, the fasting blood glucose levels decreased by 47% after 7 weeks compared with those of the control mice. According to the oral glucose tolerance test, the glucose levels recovered its baseline after 120 min in Cs-EPS-treated mice, although the blood glucose levels increased significantly after 30 min. On the other hand, the control group (not-treated) did not recovered its initial level of glucose after 120 min. Furthermore, food intake, body weight, total plasma cholesterol and triglyceride concentrations in ob/ob mice treated with Cs-EPS were significantly decreased, compared with those in control ob/ob mice. Cs-EPS treatment increased significantly the plasma insulin level and the expression of leptin mRNA in adipose tissue of Cs-EPS-treated ob/ob mice. From these results, it is demonstrated that Cs-EPS could be effective for regulating normal blood glucose levels by increasing the amounts of plasma insulin and leptin expression in ob/ob mice, indicating that this compound could be a candidate material as a dietary supplement to control hyperglycemia in patients suffering from type II diabetes.

Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi (김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명)

  • Kim Uyo-Ju;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.196-203
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    • 2006
  • Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

Biochemical Characterization of the Exopolysaccharide Purified from Laetiporus sulphureus Mycelia

  • Seo, Min-Jeong;Kang, Byoung-Won;Park, Jeong-Uck;Kim, Min-Jeong;Lee, Hye-Hyeon;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1287-1293
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    • 2011
  • The extracellular polysaccharide (EPS) was isolated from mycelial cultures of Laetiporus sulphureus var. miniatus and purified by DEAE cellulose and Sephadex G-50 column chromatography. The purified EPS (EPS-2-1) was composed of only glucose units and its molecular mass was 6.95 kDa. The chemical structure of EPS-2-1 consisted of a main chain containing ($1{\rightarrow}4$)-Glcp units with branches at the C-6 position of the chain carrying-Glcp-($1{\rightarrow}4$)-linked residues. The effect of purified EPS on immunomodulatory genes and proteins of the Bcl-2 family was observed using cultured U937 human leukemia cells. Of note, the levels of Bax and Bad proteins treated with the EPS (4 mg/ml) were approximately 23- and 18-times higher than those in non-treated cells, respectively. These results may suggest that the EPS purified from the mushroom L. sulphureus is associated with the activation of immunomodulatory mediators, Bax and Bad proteins.

Optimization of Culture Medium for the Production of an Exopolysaccharide (p-CY02) with Cryoprotective Activity by Pseudoalteromonas sp. RosPo-2 from the Antarctic Sea

  • Pilsung Kang;Sung Jin Kim;Ha Ju Park;Il Chan Kim;Se Jong Han;Joung Han Yim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1135-1145
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    • 2024
  • When cells are exposed to freezing temperatures, high concentrations of cryoprotective agents (CPA) prevent ice crystal formation, thus enhancing cell survival. However, high concentrations of CPAs can also cause cell toxicity. Exopolysaccharides (EPSs) from polar marine environments exhibit lower toxicity and display effects similar to traditional CPA. In this study, we sought to address these issues by i) selecting strains that produce EPS with novel cryoprotective activity, and ii) optimizing culture conditions for EPS production. Sixty-six bacteria producing mucous substances were isolated from the Ross Sea (Antarctic Ocean) using solid marine agar plates. Among them, Pseudoalteromonas sp. RosPo-2 was ultimately selected based on the rheological properties of the produced EPS (p-CY02). Cryoprotective activity experiments demonstrated that p-CY02 exhibited significantly cryoprotective activity at a concentration of 0.8% (w/v) on mammalian cells (HaCaT). This activity was further improved when combined with various concentrations of dimethyl sulfoxide (DMSO) compared to using DMSO alone. Moreover, the survival rate of HaCaT cells treated with 5% (v/v) DMSO and 0.8% (w/v) p-CY02 was measured at 87.9 ± 2.8% after freezing treatment. This suggests that p-CY02 may be developed as a more effective, less toxic, and novel non-permeating CPA. To enhance the production of EPS with cryoprotective activity, Response Surface Methodology (RSM) was implemented, resulting in a 1.64-fold increase in production of EPS with cryoprotective activity.

Rhizobacterial Exopolysaccharides Elicit Induced Resistance on Cucumber

  • Park, Kyung-Seok;Kloepper, Joseph W.;Ryu, Choong-Min
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1095-1100
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    • 2008
  • The role of exopolysaccharides (EPSs) from a plant growth-promoting rhizobacterium, Burkholderia gladioli IN26, on elicitation of induced systemic resistance was investigated. A purified EPS induced expression of PR-1a::GUS on tobacco and elicited induced resistance against Colletotrichum orbiculare on cucumber. The maximum level of disease protection was noted when seeds were soaked in 200 ppm of the EPS. Our results indicate that EPS from specific rhizobacteria can elicit induced resistance and suggest that bacterial EPS might be a useful elicitor of resistance under field conditions.

Fermentation Characteristics of Exopolysaccharide-Producing Lactic Acid Bacteria from Sourdough and Assessment of the Isolates for Industrial Potential

  • Jung, Seung-Won;Kim, Wang-June;Lee, Kwang-Geun;Kim, Cheol-Woo;Noh, Wan-Seob
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1266-1273
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    • 2008
  • Lactic acid bacteria (LAB) with antimicrobial activity and high exopolysaccharide (EPS) production ability isolated from sourdough were studied for their fermentation characteristics as potential new starter cultures. The values of pH, titratable acidity, and viable cell counts were $4.06{\pm}0.009-4.50{\pm}0.015,\;0.787{\pm}0.020%-1.172{\pm}0.018%,\;and\;8.78{\pm}0.08-8.98{\pm}0.06$ log CFU/ml, respectively. In order to select probiotics with a high survival rate in the gut, isolates were tested to assess resistance against the artificial gastric acid and bile juice. Viable LAB counts were significantly (p<0.05) affected by the acidity. At pH 2.0, the total declines in the initial bacterial counts were 4.52$\pm$0.07 log for S. thermophilus St-Body-1, >7.98$\pm$0.03 log for E. flavescens DU-10, >7.95$\pm$0.05 log for E. faecium DU-12, and 3.15$\pm$0.06 log for L. amylovorus DU-21. Among the strains, L. amylovorus DU-21 was the only strain that had bile tolerance under simulated gastrointestinal conditions. In order to improve EPS production by L. amylovorus DU-21, the influence of carbon source was studied. When glucose was used as a carbon source, EPS production dramatically increased to 17.19$\pm$0.28 g/l (p<0.05). The maximum cell growth (10.012$\pm$>0.012 log CFU/ml) and EPS production (18.71$\pm$0.19 g/l) were achieved when 15 g/l of glucose was employed as the carbon source.

Isolation and Characterization of Ilhizobium loti from Lotus corniculatus var. japonicus (벌노랑이로부터 Rhizobium loti 의 분리 동정 및 그 특성)

  • 소병인;이강련;윤경하;고상균;이영록
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.450-455
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    • 1992
  • Five strains of the fast-growing endosymbionts were isolated from the nodules of Lotus eorniculatus var. japonicus inhabited in Taejeon. From morphological and physiOlogical characteristics and nodulation test, the isolated strains were identified as Rhizobium loti. Compared to the control plant, both Lotus cornieulatus var. japonieus and Lotus corniculatus seedlings inoculated with the isolated strains. grew normally due to effective root nodule. The reisolated endosymbiont from the induced root nodule was confirmed identical to those of the first isolates by investigating antibiotic resistance and morphological characteristics. Three strains among the isolates. R. loti TUS I. TUS5 and TUS6 produced a ca1cof1uorbinding exopolysaccharide.

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Altered cell surface hydrophobicity of Lactobacillus paracasei KLB58 isolated from human vagina

  • Choi, Yun-Do;Oh, Eun-Taex;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.452-459
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    • 2003
  • In previous study, we have isolated Lactobacillus spp. from healthy human vagina and examined various characteristics such as antimicrobial substance production and exopolysaccharide (EPS) production. It is known that an EPS is an important factor of Lactobacillus spp. to adhere to epithelial cell. We have selected L. paracasei KLB58 having high antimicrobial activity and EPS production. In this study, we performed NTG (1-Methyl-3-nitro-1-nitrosoguanidine) mutagenesis to isolate an EPS deficienct mutant of KLB58 showing high cell surface hydrophobicity (CSll) compared to wild type strain. By monitoring the kinetics of the partition with hexadecane the EPS mutant was found to be far more hydrophobic than the wild type strain ; the CSH of the EPS mutant was 0.5-fold higher than the wild type strain.

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