• Title/Summary/Keyword: culture broth

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Partial Characterization of an Anti-Listerial Bacteriocin from Enterococcus faecium CJNU 2524

  • Yang, Jung-Mo;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.41 no.1
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    • pp.164-171
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    • 2021
  • Listeria monocytogenes is a representative foodborne pathogen and causes listeriosis. Enterococcus faecium CJNU 2524 was confirmed to produce a bacteriocin with anti-listerial activity. To establish optimal culture conditions for the production of the bacteriocin from E. faecium CJNU 2524, different media (MRS and BHI broth) and temperatures (25℃, 30℃, and 37℃) were investigated. The results showed that the optimal culture conditions were MRS broth and 25℃ or 30℃ temperatures. The crude bacteriocin was stable in a broad range of pH conditions (2.0-10.0), temperatures (60℃-100℃), and organic solvents (methanol, ethanol, acetone, acetonitrile, and chloroform). The bacteriocin activity was abolished when treated with protease but not α-amylase or lipase, indicating the proteinaceous nature of the bacteriocin. Finally, the bacteriocin showed a bactericidal mode of action against L. monocytogenes. Therefore, it can be a biopreservative candidate for controlling L. monocytogenes in dairy and meat products.

Effect of re-based Coagulants on Cell Separation Efficiency from the Culture Broth of Alcaligenes eutrophus. (Alcaligenes eutrophus의 배양액으로부터 균체 분리 효율에 미치는 철(Fe)계 응집제의 효과)

  • 류희욱;조경숙;곽종운;장용근
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.257-263
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    • 1998
  • Alcaligenes eutrophus was successfully recovered from high cell density broth by pre-treatment with Fe-based coagulants. An inorganic coagulant, Fe$_2$(SO$_4$)$_3$, and a polymerized coagulant, Ferix-3, were used. Good coagulation was observed in broad pH range of 3 to 13, the floe size was increased with increasing pH of culture broth. The optimum pH of fermentation broth for cell recovery was 10 to 13. The optimum coagulant dosages to recover cells with 95% cell recovery were increased with increasing cell concentration. Optimal coagulant dosage was lower when the polymerized coagulant was used rather than the inorganic coagulant. The coexistence of NH$_4$$\^$+/ was increased coagulant requirement, and the coagulant requirement was 0.066g Fe$_3$$\^$+//g NH$_4$$\^$+/.

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Antibacterial Activity of Water Extract of Green Tea against Pathogenic Bacteria (식중독세균에 대한 녹차 물추출물의 항균작용)

  • 박찬성
    • Food Science and Preservation
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    • v.5 no.3
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    • pp.286-291
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    • 1998
  • The sensitivity of various pathogenic bacteria(Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus 196E, Salmonella typhimurium) to the water extract of green tea was tested. Tryptic soy broth was inoculated with 10$\^$5/CPU/ml of pathogenic bacteria and incubated at 35$^{\circ}C$ for 30 hours. The extract was added at a final concentration of 0-2%(w/v) into culture broth at the mid or late exponential phase of bacteria. The growth of pathogenic bacteria was inhibited with increasing concentrations of the extract in culture broth and the late exponential phase cells were more resistant than the mid exponential phase cells. Cram positive bacteria(L. monocytogenes and S. aureus 196E) were more sensitive than Cram negative bacteria(E. coli O157:H7 and S. typhimurium). S. aureus had the highest sensitivity, followed by L monocytogenes, E. coli O157:H7. S. typhimurium was the most resistant to the water extract of green tea.

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EFFECT OF ISOLATED MICROMONOSPORA AURANTIACA ON THE FORMATION OF ARTIFICIAL PLAQUE (구강으로부터 분리한 Micromonospora aurantiaca의 인공치태 형성에 미치는 영향)

  • Yang, Kyu-Ho;Kim, Seon-Mi;Park, Jin-Kyung;Chung, Jin;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.3
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    • pp.619-626
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    • 1998
  • The critical etiologic factor in the development of dental caries is dental plaque. The main component of dental plaque is the mutan produced by Streptococcus mutans. The following results were obtained by using blue mutan to assess the factors affecting the mutan-digesting activity of Micromonospora aurantiaca isolated from oral cavity. Micromonospora aurantiaca digested more blue mutan in the minimal essential broth at pH 7.0 than at pH 5.5 or 8.5, and at $37^{\circ}C$ than at $32^{\circ}C\;or\;42^{\circ}C$. Blue mutan was similarly digested at the range of 1mM to 16mM of $CaCl_2$ and 0.1mM to 6.4 mM of $MgCl_2$, while being significantly digested at the concentration of 2.5mM of KCl. When the concentration of glucose was decreased in the minimal essential broth, the digestion of blue mutan was increased. When the culture supernatant of Micromonospora aurantiaca in the RL broth with 1% glucose or 0.5% mutan was mixed with 2 ${\times}$ BHIYS broth containing 0.5% yeast extract and 10% sucrose, the formation of artificial plaque on the orthodontic wires by Streptococcus mutans was inhibited(p<0.05). These results indicated that the production of mutanase was identified in the culture supernatant of Micromonospora aurantiaca, suppressing the formation of artificial plaque by Streptococcus mutans.

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Separation and Purification of Teicoplanin by Diaion HP-20 and Conacnavalin A Chromatography (소수성 Diaion HP-20및 친화성 Concannvalin A 크로마토그래피를 이용한 Glycopeptide계 항생제 Teicoplanin의 분리 및 정제)

  • 김창진;이재찬;박해룡;박동진;김영배
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.201-204
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    • 2003
  • Glycopeptide antibiotics, teicoplanin was purified from a mutant strain of Actinoplanes teichomyceticus ATCC31121, A. teichomyceticus MSL2211. We developed a simple procedure to separate and purify the teicoplanin from the fermentation broth. Teicoplanin was purified by two-step purification system, hydrophobic adsorption and sugar affinity chromatography in combination with HPLC analysis based on the properties of hydrophobic acyl chain and sugar moiety in teicoplanin. Teicoplanin was separated from the culture broth by Diaion HP-20 and further purified by concanavalin A affinity column chromatography. As an adsorbent resin, Diaion HP-20 in broth eliminated toxic effects on growth, reduced feedback repression of teicoplanin production, and assisted In rapid recovery of teicoplanin. The teicoplanin displayed the final yield of 80% and 95% of purity.

Viability of eggs, filariform larvae and adults of Stronglyloides venezuelensis (Nematoda: Strongyloidea) maintained in vitro (베네수엘라분선충 (Strongvloides venezuelensis)의 충란, 감염자충 및 성충의 실험관 내 배양)

  • ;M.
    • Parasites, Hosts and Diseases
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    • v.36 no.2
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    • pp.99-108
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    • 1998
  • The present study was performed to check the viability of eggs, filariform larvae and adults of Strongvloines venezueLensis exposed to various conditions for an in vitro maintenance. The eggs in the feces remained viable for about 25 days at $4^{\circ}C$ and 15 days at room temperature. However, the isolated eggs in sterile saline lost their viability within 24 hr at $4^{\circ}C$. The eggs in morula stage were very sensitive to air drying and rapidly lost their viability (=12 hrs. Filariform larvae survived for a maximum period of 45 days in fecal suspension and 28 days in 0.12% nutrient broth in polyvinyl culture bags maintained at $20^{\circ}C$. On the other hand, those isolated from nutrient broth cultures survived for a maximum period of 32 days in tap water and 22 days in sterile saline at $20^{\circ}C$. The mature adult worms obtained from experimentally infected rats survived maximally for 9 days in serum supplemented (10% rat-serum) 0.12% nutrient broth and 4 days in serum free nutrient broth at $37^{\circ}C$ while the culture media were changed at an alternate day. The adult female worms deposited fertile eggs in serum supplemented and serum free nutrient broth cultures, however, the hatched larvae (Ll) were not able to develop to the filariform stage in the culture media and found to die within 24 hr of maintenance. The present findings on an in vitro maintenance of different stages of 5. uenezueLetis may provide useful information for biological and biochemical studies with Strongyloines species. Key words: Strongvloides venezuelensis. viability in vitro maintenance, free-living filariform larvae (L3), embryonation of eggs

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Characteristics of Rahnella aquatilis Strain AY2000 for an Anti-Yeast Substance Production (항효모성 물질 생산을 위한 Rahnella aquatilis AY2000 균주의 생육특성)

  • Kang, Min-Jung;Lee, Bok-Kyu;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.215-220
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    • 2008
  • Rahnella aquatilis AY2000 has an unique characteristic which produces an anti-yeast substance (AYS). The AYS of the strain AY2000 was always secreted on agar plate, however, its activity in liquid culture was labile upon storage of the medium. In this paper, cultural conditions of the strain AY2000 for the AYS production were investigated in liquid culture, and minimal inhibitory concentration (MIC) against Saccharomyces cerevisiae was determined for the AYS activity. MIC of the AYS cultured in PYG broth at $^25{\circ}C$ for 24 hr was $23.5{\mu}g/mL$, however, that in MYCS (pH 5.5) broth at the same condition was $15.5{\mu}g/mL$. The activity of the AYS had increased rather in MYCS broth excluded $NH_4$-citrate than in the same broth contained $NH_4$-citrate, and MIC of the AYS produced in MYCS broth without $NH_4$-citrate was $15.5{\mu}g/mL$. When the strain AY2000 was maintained in MYCS broth without $NH_4$-citrate but added $100{\mu}M$ $FeCl_3$, the activity of the AYS had increased and its MIC was $7.8{\mu}g/mL$. MIC of the AYS was $7.8{\mu}g/mL$ after the strain AY2000 was cultured in MYCS broth containing $100{\mu}M$ $FeCl_3$ without $NH_4$-citrate, however, its MIC was $31.3{\mu}g/mL$ after 48-60 hr culture in the same broth.

Production of Indigo and Indirubin by Escherichia coli Containing a Phenol Hydroxylase Gene of Bacillus stearothermophilus

  • Kim, In-Cheol;Chang, Hae-Choon;Oriel, Patrick
    • Journal of Microbiology and Biotechnology
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    • v.7 no.3
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    • pp.197-199
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    • 1997
  • Escherichia coli recombinants containing the cloned phenol hydroxylase gene of Bacillus stearothermophilus BR219 were shown to produce both indigo and its structural isomer indirubin during culture on LB broth. The ratio of indirubin/indigo was highest under conditions of prolonged culture and reduced culture oxygenation.

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In Vitro Proliferation Model of Helicobacter pylori Required for Large-Scale Cultivation

  • Oh, Heung-Il;Lee, Heung-Shick;Kim, Kyung-Hyun;Paek, Se-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.367-374
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    • 2000
  • The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

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Development of W/O/W Multiple Emulsion Formulation Containing Burkholderia gladioli

  • KIM, HWA-JIN;CHO, YOUNG-HEE;BAE, EUN-KYUNG;SHIN, TAEK-SU;CHOI, SUNG-WON;CHOI, KEE-HYUN;PARK, JI-YONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.29-34
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    • 2005
  • W/O/W (water-in-oil-in-water) type multiple emulsion was applied to improve the storage stability of an antagonistic microorganism, Burkholderia gladioli. Encapsulation of microorganism into a W/O/W emulsion was conducted by using a two-step emulsification method. W/O/W emulsion was prepared by the incorporation of B. gladioli into rapeseed oil and the addition of polyglycerin polyriconolate (PGPR) and castor oil polyoxyethylene (COG 25) as the primary and secondary emulsifier, respectively. Microcrystalline cellulose was used as an emulsion stabilizer. To evaluate the usefulness of W/O/W emulsion formulation as a microbial pesticide for controlling the bacterial wilt pathogen (Ralstonia solanacearum), the storage stability and antagonistic activity of emulsion formulation were tested in vitro. The storage stability test revealed that the viability of formulated cells in emulsion was higher than that of unformulated cells in culture broth. At $4^{\circ}C$, the viabilities of formulated cells and unformulated cells at the end of 20 weeks decreased to about 2 and 5 log cycles, respectively. At $37^{\circ}C$, the viability of formulated cells decreased to only 2 log cycles at the end of storage. On the other hand, the viable cells in culture broth were not detected after 13 weeks. In activity test, formulated cells in emulsion were more effective in inhibiting the growth of pathogen than unformulated cells in culture broth. Unformulated cells completely lost their antagonistic activity during storage under similar conditions. The W/O/W multiple emulsion formulation was shown to be useful as the novel liquid formulation for biological control.